TW201307345A - Crystalline (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]phthalazin-3(7H)-one tosylate salt - Google Patents

Abstract

Provided herein are (8S, 9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1, 2, 4-triazol-5-yl)-8, 9-dihydro-2H-pyrido[4, 3, 2-de]phthalazin-3(7H)-one tosylate salt forms, including crystalline forms, and methods of their preparation. Pharmaceutical compositions comprising a (8S, 9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1, 2, 4-triazol-5-yl)-8, 9-dihydro-2H-pyrido[4, 3, 2-de]phthalazin-3(7H)-one tosylate salt are also provided, as are methods of using (8S, 9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1, 2, 4-triazol-5-yl)-8, 9-dihydro-2H-pyrido[4, 3, 2-de]phthalazin-3(7H)-one tosylate salt to treat a disease or condition, such as a cancer.

Description

Crystalline (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9- Dihydro-2H-pyrido[4,3,2-de]indole-3(7H)-one tosylate

This application relates to (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5-yl )-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3( 7H )-ketotosylate form, in crystalline form; preparation of ( 8S , 9R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl- 1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 a method of -3( 7H )-ketone and its tosylate form; and including crystalline ( 8S , 9R )-5-fluoro-8-(4-fluorophenyl)-9-(1-A Base-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 A pharmaceutical composition in the form of -3( 7H )-ketotosylate.

Cross-reference to related applications

The present application claims priority to U.S. Provisional Application Serial No. 61/405,476, filed on Jan. 21, 2010, the content of which is hereby incorporated by reference.

Enzymes in the poly(ADP-ribose) polymerase (PARP) protein family are involved in a variety of cellular functions, including helping to repair DNA damage. Inhibition of PARP activity is a promising therapeutic approach to the treatment of certain cancers. A variety of PARP inhibitors are in clinical and preclinical stages as candidates for the treatment of ovarian, breast, colorectal, prostate and other cancers. One type of PARP inhibitor disclosed in US Publication No. 2010/0035883 comprises (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2, 4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-one. Preclinical studies have shown that this compound is available for some cancer patients.

U.S. Publication No. 2010/0035883 provides 5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8, 9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3 (7 H) - Synthesis and Its -one (8 S, 9 R) enantiomer of the resolved chiral. See, for example, US 2010/0035883, Examples 94 and 155, which are hereby incorporated by reference in their entirety for all purposes. Other methods of preparing such compounds are described in WO 2011/097602. For the like (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)- 8,9-Dihydro-2 H -pyrido[4,3,2-de]呔 Candidate drugs such as -3(7 H )-ketone are transferred to viable pharmaceutical products. It is important to know whether the candidate drug has a polymorphic form and that the form is produced, transported, stored and used before mass production, transportation, storage and use. Relative stability and interconversion under conditions that may be encountered during preparation. The ability to control and generate stable polymorphs with robust processes can be key to regulatory approval and sales. The industry seeks high purity 5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro- 2 H -pyrido[4,3,2- de ]呔 A large-scale production method of -3( 7H )-ketone, which is subject to the fact that the parameters affecting the stability of the compound have not been known and the polymorphic form of the compound has not been obtained.

In one aspect, (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5-yl is provided herein. )-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketotosylate. In some embodiments, the tosylate salt is in crystalline form. In some embodiments, provided herein is (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole- 5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 A solid form of the tosylate salt of -3( 7H )-ketone, comprising a crystalline form, an amorphous form, or a mixture thereof.

In another aspect, the invention provides (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H in the form of the free base ("free base"). -1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketone method.

In another aspect, the preparation of (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5- ,8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketone tosylate method. In certain embodiments, the tosylate salt is prepared from a suspension in tetrahydrofuran (THF).

In another aspect, the application provides (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1, as described herein, 2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketotosylate or a pharmaceutical composition thereof in solid form.

In another aspect, the application provides (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4 -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 A method of treating cancer by -3 ( 7H )-ketotosylate (in another example, a crystalline form thereof) or a composition thereof.

(8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9 -dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketone is a promising candidate for the treatment of, for example, the following cancers: leukemia (including acute myeloid leukemia, chronic lymphocytic leukemia), myelodysplastic syndrome, colon cancer, EBV- Related tumors (including Burkitt's lymphoma, nasopharyngeal carcinoma, lymphoma of AIDS patients, smooth muscle tumors of AIDS patients, Hodgkin's disease, non-Hodgkin's lymphomas -Hodgkin's lymphoma), lymphoproliferative disease in immunosuppressed patients, leiomyosarcoma in immunosuppressed patients, EBV+gastric cancer, EBV+ breast cancer, T-cell lymphoma), endometrial cancer (including cancer and sarcoma), gastrointestinal matrix tumor, Glioma, glioblastoma, lymphoma (including mantle cell lymphoma), melanoma, breast cancer (including metastatic breast cancer, BRCA-positive breast cancer, and BRCA-negative breast cancer), ovarian cancer (including advanced ovarian cancer, highly differentiated serum) Ovarian cancer, platinum-sensitive ovarian cancer, platinum-resistant ovarian cancer, platinum-refractory ovarian cancer, and BRCA-negative ovarian cancer), cervical cancer, pancreatic cancer (including BRCA-negative pancreatic cancer), peritoneal cancer, Adenocarcinoma (including BRCA-negative prostate cancer, metastatic prostate cancer, and castration-resistant prostate cancer), hereditary non-polyposis colon cancer (HNPCC), lung cancer (including non-small cell lung cancer, small cell lung cancer), colorectal cancer , uterine carcinosarcoma, solid tumors (such as bladder, intestine, brain, breast, endometrium, heart, kidney, lung, non-pancreatic endocrine (thyroid) cancer and solid tumor of the head and neck) and blood tumors.

Provided herein are methods of producing a compound, a form of the tosylate salt comprising the compound. The method provided herein is the previously reported free form of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1, 2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 (7 H) - one Synthesis of (e.g., see US 2010/0035883) compared to provide (e.g.) improvement of the product recovery and / or fewer processing steps, and is suitable for mass production of the compound. In one embodiment, as shown in the examples below, (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-) is prepared in crystalline form. 1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The stability of the -3( 7H )-ketotosylate exceeds other solid forms associated with the free base and other salts of the compound, or has other advantageous properties. The examples provided herein demonstrate that certain different solvents can be used to form ( 8S , 9R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1, 2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 ( 7H )-keto tosylate polymorph.

the term

A number of terms are defined below to aid in understanding the disclosure described herein. Generally, the nomenclature used herein and the laboratory procedures for organic chemistry, medical chemistry, and pharmacology described herein are well known and commonly employed in the art. All technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art of the present disclosure, unless otherwise defined.

abbreviation

As used herein, when referring to "(8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5 -yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketone" or its formula,

Unless otherwise stated or explicitly indicated in the context of the reference, it refers to the free base form of the compound.

It should be understood that (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)- 8,9-Dihydro-2 H -pyrido[4,3,2- de ]呔 -3( 7H )-ketotosylate includes (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2 ,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 a cation of -3( 7H )-ketone (e.g., in one embodiment, protonated at one atomic position, or in other embodiments, protonated at more than one atomic position) and p-toluenesulfonic acid An anion wherein the anion is referred to herein as "tosylate." In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 -yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 The solid form of the -3( 7H )-ketotosylate can include a molar ratio of cation to anion of about 1:1. In certain embodiments, the molar ratio of cation to anion in the solid salt can be about 1:1.33, about 1:1.5, or about 1:2.

The use of the terms "about" and "about" when used in combination with the ingredients, dosages, or percentages of ingredients of the compositions or dosage forms are intended to be recognized by those skilled in the art, unless otherwise specified. Dosage, amount or percentage by weight of the pharmacological effect equivalent to the pharmacological effect obtained from the specified dose, amount or percentage by weight may be provided. Specifically, as used herein, the terms "about" and "about" encompass 15%, 10%, 5%, 4%, 3%, 2%, 1% or 0.5 of the specified dose, amount or weight percentage. Dosage, amount or percentage by weight of %.

The terms "about" and "approximately", as used herein, are used in connection with the <RTI ID=0.0></RTI></RTI><RTIgt; Temperature or temperature range of solvation or glass transition; mass change, such as mass with temperature or humidity; solvent or water content expressed, for example, by mass or percentage; or peak position, such as ¹³ C NMR, DSC, TGA, and The peak position in the XRPD analysis is indicated when used, and the value or range of values may deviate from the extent to which the skilled artisan would be reasonable and still state the particular solid form. In particular, the terms "about" and "approximately" are used in this context to indicate that the value or range of values may differ from the recited value or range of values by 5%, 4%, 3%, 2%, 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2% or 0.1% and this particular solid form is still set forth.

The term "amorphous" or "amorphous form" is intended to mean that the substance, component or product in question is substantially non-crystalline, as determined by, for example, XRPD, or the substance or component in question Or the product is, for example, not birefringent when viewed under a microscope. In certain embodiments, a sample comprising an amorphous form of a substance may be substantially free of other amorphous forms and/or crystalline forms.

The term "crystalline form" or "crystal form" means a crystalline solid form of a chemical compound, including but not limited to a single component or multi-component crystal form, for example, a polymorph of a compound; or a solvent combination of a compound , hydrate, clathrate, co-crystal, salt or polymorph thereof. The term "crystalline form" and related terms herein, refers to various crystalline forms of a given substance, including but not limited to polymorphs, solvates, hydrates, co-crystals and other molecular complexes, and salts. , solvates of salts, hydrates of salts, other molecular complexes of salts and polymorphs thereof. The crystalline form of the material can be obtained by a variety of methods known in the art. Such methods include, but are not limited to, melt recrystallization, melt cooling, solvent recrystallization, recrystallization in a confined space (eg, in a nanopore or capillary), on a surface or template (eg, at Recrystallization on the polymer, recrystallization, desolvation, dehydration, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, sublimation, grinding and solvent micromilling in the presence of additives (eg, co-crystal counter molecules).

Techniques for characterizing crystalline forms and amorphous forms include, but are not limited to, TGA, DSC, XRPD, single crystal X-ray diffraction, vibrational spectroscopy (eg, IR and Raman spectroscopy), solid state NMR, Optical microscopy, high temperature stage optical microscopy, SEM, electronic crystallography and quantitative analysis, PSA, surface area analysis, solubility studies, and dissolution studies.

As used herein and unless otherwise indicated, the term "hydrate" means a compound or a salt thereof, further comprising a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces. As used herein and unless otherwise indicated, the term "solvate" means a solvate formed by combining one or more solvent molecules with a compound provided herein or a salt thereof. The term "solvate" encompasses hydrates (eg, hemihydrate, monohydrate, dihydrate, trihydrate, tetrahydrate, and the like). (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro -2H-pyrido[4,3,2-de]呔 The solvate (e.g., hydrate) of -3(7H)-ketone may be crystalline or amorphous.

The term "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid filler or solid filler, diluent, solvent or encapsulating material. In one embodiment, the meaning of "pharmaceutically acceptable" for each component is that it is compatible with the other ingredients of the pharmaceutical formulation and is suitable for contact with tissues and organs of humans and animals without undue toxicity or irritation. , allergic reactions, immunogenicity or other problems or complications, commensurate with a reasonable benefit/risk ratio. See, for example, Remington: The Science and Practice of Pharmacy, 21st Ed .; Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 6th Edition; Rowe et al., ed .; The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd edition ; Editing by Ash and Ash; Gower Publishing: 2007; Pharmaceutical Preformulation and Formulation, 2nd edition; Gibson Editor; CRC Press LLC: Boca Raton, FL, 2009.

The term "polymorph" or "polymorphic form" refers to one of two or more crystal forms comprising the same molecule or ion. Different polymorphs may have different physical properties such as melting temperature, heat of fusion, solubility, dissolution rate, and/or vibrational spectrum due to the arrangement or conformation of molecules or ions in the crystal lattice. The differences in physical properties exhibited by polymorphs can affect pharmaceutical parameters such as storage stability, compressibility, density (important in formulation and product manufacture), and dissolution rate (an important factor in bioavailability). The difference in stability can be a change in chemical reactivity (eg, differential oxidation, which causes the dosage form to fade more rapidly when including one polymorph than when including another polymorph), mechanical changes (eg, in storage, with The kinetically favorable polymorph is converted to a thermodynamically more stable polymorph, tablet pulverization) or both (for example, a tablet of a polymorph is more susceptible to decomposition under high humidity). Due to differences in solubility/dissolution, in extreme cases, some polymorphic transformations may result in insufficient potency or, in the other extreme, toxicity. In addition, the physical properties of the crystalline form may be important in the treatment; for example, a polymorph may be more susceptible to solvate formation or may be difficult to filter and wash out impurities (eg, between polymorphs, particle shapes and particles) The diameter distribution may be different).

As used herein and unless otherwise indicated, the term "stereoisomerically pure" means that the composition includes one stereoisomer of the compound and is substantially free of other stereoisomers of the compound. In certain embodiments, provided herein are stereoisomerically pure (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1, 2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 a -3( 7H )-one which is substantially free of, for example, (8R,9S)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1 ,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3 ( 7H )-one other stereoisomer of ketone. In certain embodiments, the stereoisomerically pure compound comprises greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, greater than about 90% by weight of the compound. a stereoisomer and less than about 10% by weight of other stereoisomers of the compound, greater than about 95% by weight of one stereoisomer of the compound, and less than about 5% by weight of other stereoisomers of the compound, greater than One stereoisomer of about 97% by weight of the compound and less than about 3% by weight of other stereoisomers or more than about 99% by weight of one stereoisomer of the compound and less than about 1% by weight of other stereoisomers of the compound Structure. In certain embodiments, the term "stereoisomerically pure" of (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1 ,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 The -3( 7H )-ketone means that the compound is made up of about 100% by weight of this particular stereoisomer. The above percentages are based on the total amount of the combined stereoisomers of the compounds.

As used herein, "pure" (ie, substantially free of other crystalline or amorphous forms) crystalline or amorphous forms contain less than about 10% by weight of one or more other crystalline or amorphous forms. , less than about 5% by weight of one or more other crystalline or amorphous forms, less than about 3% by weight of one or more other crystalline or amorphous forms, less than about 1% by weight of one or more other crystalline forms or Amorphous form or less than about 0.5% by weight of one or more other crystalline or amorphous forms. In some cases, as used herein, "substantially pure" (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H - 1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3(7 H )-one or a salt or solvate thereof may mean no other chemical compound, for example, in the preparation of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)- 9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 Unreacted precursors and by-products that may be present in the -3( 7H )-ketone process. In other cases, as used herein, (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-tri Zyrid-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 A "substantially pure" solid form of -3( 7H )-one or a salt or solvate thereof (eg, crystalline form or amorphous form) may mean that (8 S , 9 R )-5 is absent. -Fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyridine [4,3,2- de ]呔 -3( 7H )-one or other solid form of its salt or solvate. Thus, in certain embodiments, "substantially pure" (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1, 2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3(7 H )-one may comprise less than about 10%, 5%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or 0.1% by weight of the compound One or more other crystalline forms and amorphous forms and/or other chemical compounds. In certain embodiments, a substantially pure solid form is substantially free of one or more other specific crystalline forms, amorphous forms, and/or other chemical compounds.

The term "individual" refers to an animal including, but not limited to, a primate (eg, a human), a monkey, a cow, a pig, a sheep, a goat, a horse, a dog, a cat, a rabbit, a rat, or a mouse. As used herein, the terms "individual" and "patient" are used interchangeably when referring to, for example, a mammalian individual (eg, a human).

The term "treat, treating, and treating" is intended to include alleviating or eliminating a condition, disease, or condition, or one or more symptoms associated with the condition, disease, or condition; or slowing down the disease, condition, or condition, or one or more of its symptoms. Progress, proliferation or deterioration. In general, the beneficial effects of an individual from a therapeutic agent do not completely cure the disease, condition or condition.

The term "therapeutically effective amount" is intended to include an amount of the compound which, when administered, is sufficient to prevent or reduce one or more of the symptoms of the condition, disease or condition being treated to a certain extent. The term "therapeutically effective amount" also refers to a compound sought by a researcher, veterinarian, physician or clinician sufficient to elicit a biological or medical response to, for example, inhibit PARP activity in vivo, inhibit cancer cell growth and/or proliferation and/or Or reduce the amount of cancer cells.

As used herein, the term "vol" means the weight/volume ratio of a solid reactant to a liquid solvent. For example, the presence of 250 g of solid material in 10 vol of solvent means that the material is dissolved in 10 x 250 mL or 2.5 L of solvent.

Example

In one aspect, (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5-yl is provided herein. )-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketotosylate.

In some embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-tri" provided herein Zyrid-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketotosylate is in crystalline form. In some embodiments, the crystalline form is not solvated. In other embodiments, the crystalline form is a solvate. For example, the crystalline solvate form can be a hydrate. In other embodiments, (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5 is provided herein. -yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 The -3(7H)-ketotosylate salt is in an amorphous form. In other embodiments, provided herein is (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole- 5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 a solid form (e.g., a crystalline form, an amorphous form, or a mixture of forms) of -3( 7H )-one or a salt or solvate thereof (e.g., a salt provided elsewhere herein). In one embodiment, provided herein is (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole- 5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 a crystalline form of -3( 7H )-one or a salt or solvate thereof. In one embodiment, provided herein is (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole- 5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 An amorphous form of -3( 7H )-one or a salt or solvate thereof.

In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4- provided herein Triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The crystalline form of -3( 7H )-ketotosylate is substantially pure. For example, in various embodiments, the crystalline tosylate salt is at least about 90%, at least about 95%, at least about 97%, at least about 98%, at least about 99%, at least, by weight of the single crystal form. About 99.2%, at least about 99.5%, at least about 99.6%, at least about 99.7%, or at least about 99.8%, the balance of the total weight may be in other crystalline or amorphous forms and/or other compounds. In one embodiment, the crystalline tosylate salt is substantially in a single component crystalline form or a single polymorph. In another embodiment, the crystalline tosylate salt is a multicomponent crystalline form comprising (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-A Base-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 a first crystalline form of -3( 7H )-ketone and at least one other crystalline and/or amorphous form. In some or any embodiments, the crystalline form is substantially free of (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1, 2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 Amorphous form of -3( 7H )-ketone.

Unless otherwise specified, when the XRPD peak is represented by the 2θ angle, it is understood that copper Kα1 radiation is used. In some embodiments, the 2 theta angle values provided herein can vary within the range of ± 0.2 ° θ and still illustrate the same XRPD peak.

In ¹³ C solid-state NMR, the peak position can be changed by factors such as: signal-to-noise ratio, peak width, temperature, rotation rate, decoupling efficiency, magic angle setting, data processing program and parameters, and software peak extraction algorithm. . In addition, the peak position is related to the chemical shift reference procedure. Several different chemical shift reference standards can be used and the same results are not necessarily obtained. This can result in a difference in peak position by a few ppm. However, if different reference standards are used or if the analyst uses different values for the reference peak positions of the same standard, then generally all peak positions will have regular changes in the same direction. In some embodiments, in ¹³ C solid state NMR, the ppm values provided herein can vary from about ± 0.2 ppm while still illustrating the same peak.

In certain embodiments, (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole- 5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 a crystalline salt of a -3( 7H )-ketone having one or more XRPD patterns (eg, 1, 2, 3, 4, 5, 6, 7, 8, 9) 10 or more than 10; or at least 3, at least 4, at least 5, at least 6, or at least 7) selected from the group consisting of any of Tables 9, 11, 13, 15, 17 and 25 -value( ) indicates the characteristic peak of the peak. In another embodiment, the crystalline salt tosylate salt has one or more (eg, 1, 2, 3, 4, 5, 6, 7, 8, 9 , 10 or more than 10; or at least 3, at least 4, at least 5, at least 6, or at least 7) selected from any one of Tables 9, 11, 13, 15, 17 and 25 The XRPD peak of the peak of the 2θ angle. In certain embodiments, the crystalline tosylate salt has an XRPD pattern substantially as provided in Figures 1, 6 or 8.

In certain embodiments, the XRPD pattern of crystalline tosylate salt provided herein comprises one or more (eg, 1, 2, 3, 4, 5, 6, 7, 8 , 9, 10 or more than 10; or at least 3, at least 4, at least 5, at least 6, or at least 7) selected from the 2θ angles or d values as provided in Table 15 ( The XRPD peak of the peak. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.79, 5.86, 4.90, 4.42, 4.35, 3.93, and 3.70. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.79, 5.86, 4.90, 4.65, 4.42, 4.35, 4.13, 3.93, and 3.70. In certain embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), such peaks From d value ( ) are peaks of about 11.79, 5.86, 4.98, 4.90, 4.79, 4.65, 4.42, 4.35, 4.13, 3.93, 3.70, and 3.58. In certain embodiments, the XRPD pattern of crystalline tosylate comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), such peaks From d value ( ) includes peaks of 11.79, 7.07, 6.13, 5.86, 5.10, 4.98, 4.90, 4.79, 4.65, 4.42, 4.35, 4.13, 4.08, 3.93, 3.85, 3.70, 3.58, 3.31, and 2.99. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, they are peaks of about 7.49, 15.10, 18.10, 20.06, 20.40, 22.61, and 24.01. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.49, 15.10, 18.10, 19.08, 20.06, 20.40, 21.49, 22.61, and 24.01. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.49, 15.10, 17.78, 18.10, 18.49, 19.08, 20.06, 20.40, 21.49, 22.61, 24.01, and 24.84. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles are peaks of about 7.49, 12.50, 14.44, 15.10, 17.38, 17.78, 18.10, 18.49, 19.08, 20.06, 20.40, 21.49, 21.76, 22.61, 23.05, 24.01, 24.84, 26.93 and 29.82.

In certain embodiments, the XRPD pattern of crystalline tosylate comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), such peaks Selected from the 2θ angle or d value as provided in Table 15 ( The peak. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 5.9, 4.9, 4.42, 4.35, 3.9, and 3.7. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 5.9, 4.9, 4.7, 4.42, 4.35, 4.1, 3.9, and 3.7. In certain embodiments, the XRPD pattern includes one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks) selected from d values ( ) includes peaks of about 11.8, 5.9, 5.0, 4.9, 4.8, 4.7, 4.42, 4.35, 4.1, 3.9, 3.70, and 3.58. In certain embodiments, the XRPD pattern comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks) selected from d values ( ) includes peaks of about 11.8, 7.1, 6.1, 5.9, 5.1, 5.0, 4.9, 4.8, 4.7, 4.42, 4.35, 4.1, 4.1, 3.9, 3.9, 3.7, 3.6, 3.3, and 3.0. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.5, 15.1, 18.1, 20.1, 20.4, 22.6, and 24.0. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.5, 15.1, 18.1, 19.1, 20.1, 20.4, 21.5, 22.6, and 24.0. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, they are peaks of about 7.5, 15.1, 17.8, 18.1, 18.5, 19.1, 20.1, 20.4, 21.5, 22.6, 24.0, and 24.8. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles are peaks of about 7.5, 12.5, 14.4, 15.1, 17.4, 17.8, 18.1, 18.5, 19.1, 20.1, 20.4, 21.5, 21.8, 22.6, 23.1, 24.0, 24.8, 26.9 and 29.8.

In certain embodiments, the crystalline tosylate salt comprises a 2 theta angle or d value as provided in Table 9 ( ) XRPD peak. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.42, 15.01, 17.70, 18.01, 18.47, 18.98, 19.98, 20.33, 21.41, 22.58, 23.95, and 24.76. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.42, 15.01, 18.01, 19.98, 20.33, 22.58, and 23.95. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.42, 15.01, 17.70, 18.01, 19.98, 20.33, 21.41, 22.58, 23.95, and 24.76. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.90, 5.90, 5.01, 4.92, 4.44, 4.37, 4.15, 3.93, 3.71, and 3.59. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.90, 5.90, 5.01, 4.92, 4.80, 4.67, 4.44, 4.37, 4.15, 3.93, 3.71, and 3.59. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) is the peak of about 11.90, 5.90, 4.92, 4.44, 4.37, 3.93 and 3.71.

In certain embodiments, the XRPD pattern of crystalline tosylate comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), such peaks Selected from the 2θ angle or d value as provided in Table 9 ( The peak. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, they are peaks of about 7.4, 15.0, 17.7, 18.0, 18.5, 19.0, 20.0, 20.3, 21.4, 22.6, 24.0, and 24.8. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, they are peaks of about 7.4, 15.0, 18.0, 20.0, 20.3, 22.6, and 24.0. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.4, 15.0, 17.7, 18.0, 20.0, 20.3, 21.4, 22.6, 24.0, and 24.8. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.9, 5.9, 5.0, 4.9, 4.44, 4.37, 4.1, 3.9, 3.7, and 3.6. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.9, 5.9, 5.0, 4.9, 4.8, 4.7, 4.44, 4.37, 4.1, 3.9, 3.7, and 3.6. In some embodiments, the crystalline tosylate salt comprises a d value ( ) are XRPD peaks of about 11.9, 5.9, 4.9, 4.44, 4.37, 3.9, and 3.7.

In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected Since having a 2θ angle or d value as provided in Table 11 ( The peak. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.42, 15.02, 17.38, 17.74, 18.03, 18.54, 19.02, 20.08, 20.39, 21.44, 22.63, 24.00, and 24.83. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.42, 15.02, 18.03, 20.08, 20.39, 22.63, and 24.00. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of approximately 11.91, 5.89, 5.10, 5.00, 4.92, 4.78, 4.66, 4.42, 4.35, 4.14, 3.93, 3.71 and 3.58. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) is the peak of about 11.91, 5.89, 4.92, 4.42, 4.35, 3.93 and 3.71.

In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected Since having a 2θ angle or d value as provided in Table 11 ( The peak. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.4, 15.0, 17.3, 17.7, 18.0, 18.5, 19.0, 20.1, 20.4, 21.4, 22.6, 24.0, and 24.8. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, they are peaks of about 7.4, 15.0, 18.0, 20.1, 20.4, 22.6, and 24.0. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.9, 5.9, 5.1, 5.0, 4.9, 4.8, 4.7, 4.42, 4.35, 4.1, 3.9, 3.7, and 3.6. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.9, 5.9, 4.9, 4.42, 4.35, 3.9, and 3.7.

In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected Since having a 2θ angle or d value as provided in Table 13 ( The peak. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.46, 12.47, 14.45, 15.09, 17.40, 17.74, 18.11, 18.53, 19.05, 20.09, 20.43, 21.46, 22.63, 23.10, 24.03, 24.85, and 26.96. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.46, 14.45, 15.09, 17.74, 18.11, 18.53, 19.05, 20.09, 20.43, 21.46, 22.63, 24.03, 24.85, and 26.96. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, they are peaks of about 7.46, 15.09, 18.11, 20.09, 20.43, 22.63, and 24.03. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.84, 7.09, 6.13, 5.87, 5.09, 5.00, 4.89, 4.78, 4, 66, 4.42, 4.34, 4.13, 4.08, 3.93, 3.85, 3.70, 3.58, 3.30, 2.99, and 2.86. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.84, 6.13, 5.87, 5.09, 5.00, 4.89, 4.78, 4.42, 4.34, 4.13, 3.93, 3.70, 3.58, and 3.30. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) is the peak of about 11.84, 5.87, 4.89, 4.42, 4.34, 3.93 and 3.70.

In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected Since having a 2θ angle or d value as provided in Table 13 ( The peak. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.5, 12.5, 14.5, 15.1, 17.4, 17.7, 18.1, 18.5, 19.0, 20.1, 20.4, 21.5, 22.6, 23.1, 24.0, 24.9, and 27.0. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.5, 14.5, 15.1, 17.7, 18.1, 18.5, 19.0, 20.1, 20.4, 21.5, 22.6, 24.0, 24.9, and 27.0. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.5, 15.1, 18.1, 20.1, 20.4, 22.6, and 24.0. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 7.1, 6.1, 5.9, 5.1, 5.0, 4.9, 4.8, 4.7, 4.4, 4.3, 4.13, 4.08, 3.9, 3.8, 3.7, 3.6, 3.3, 3.0, and 2.9. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 6.1, 5.9, 5.0, 4.9, 4.8, 4.7, 4.4, 4.3, 4.1, 3.9, 3.7, 3.6, and 3.3. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 5.9, 4.9, 4.4, 4.3, 3.9, and 3.7.

In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected Since having a 2θ angle or d value as provided in Table 17 ( The peak. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From 2θ angles are about 7.50, 12.51, 14.48, 15.12, 17.38, 17.78, 18.17, 18.58, 19.11, 20.09, 20.54, 21.54, 21.86, 22.65, 23.19, 24.08, 24.86, 26.98, 29.97, 30.44, 30.84, 32.07, 32.49 And the peak of 37.56. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles are peaks of about 7.50, 15.12, 17.38, 17.78, 18.17, 18.58, 19.11, 20.09, 20.54, 21.54, 21.86, 22.65, 23.19, 24.08, 24.86 and 26.98. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, they are peaks of about 7.50, 15.12, 18.17, 20.09, 20.54, 22.65, and 24.08. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of approximately 11.78, 7.07, 6.11, 5.85, 5.10, 4.98, 4.88, 4.77, 4.64, 4.42, 4.32, 4.12, 4.06, 3.92, 3.83, 3.69, 3.57, 3.30, 2.98, 2.93, 2.78, 2.75, 2.39 . In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.78, 5.85, 5.10, 4.98, 4.88, 4.77, 4.64, 4.42, 4.32, 4.12, 4.06, 3.92, 3.83, 3.69, 3.57, and 3.30. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) is the peak of about 11.78, 5.85, 4.88, 4.42, 4.32, 3.92 and 3.69.

In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected Since having a 2θ angle or d value as provided in Table 17 ( The peak. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From 2θ angles are about 7.5, 12.5, 14.5, 15.1, 17.4, 17.8, 18.2, 18.6, 19.1, 20.1, 20.5, 21.5, 21.9, 22.6, 23.2, 24.1, 24.9, 27.0, 30.0, 30.4, 30.8, 32.1, 32.5 And the peak of 37.6. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.5, 15.1, 17.4, 17.8, 18.2, 18.6, 19.1, 20.1, 20.5, 21.5, 21.9, 22.6, 23.2, 24.1, 24.9, and 27.0. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, they are peaks of about 7.5, 15.1, 18.2, 20.1, 20.5, 22.6, and 24.1. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) is the peak of about 11.8, 7.1, 6.1, 5.9, 5.1, 5.0, 4.9, 4.8, 4.6, 4.4, 4.3, 4.12, 4.06, 3.9, 3.8, 3.7, 3.6, 3.3, 3.0, 2.9, 2.78, 2.75, 2.4 . In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 5.9, 5.1, 5.0, 4.9, 4.8, 4.6, 4.4, 4.3, 4.12, 4.06, 3.9, 3.8, 3.7, 3.6, and 3.3. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 5.9, 4.9, 4.4, 4.3, 3.9, and 3.7.

In some embodiments, the crystalline tosylate salt has an XRPD pattern substantially as provided in Figure 6. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the peak with the 2 theta angle as provided in Table 25. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.51, 14.47, 15.14, 17.41, 18.12, 18.53, 19.07, 20.09, 20.46, 21.48, 21.81, 24.05, 24.83, and 29.81 (± about 0.2). In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected The peaks at about 2θ are about 7.51, 14.47, 15.14, 20.09, 21.48, and 24.05 (± about 0.2). In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.51, 15.14, 18.12, 20.09, 20.46, 22.65, and 24.05 (± about 0.2). In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected The peaks at about 2θ are about 7.51, 20.09, and 24.05 (± about 0.2). In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles are peaks of about 7.5, 14.5, 15.1, 17.4, 18.1, 18.5, 19.1, 20.1, 20.46, 21.48, 21.8, 24.1, 24.8, and 29.8 (± about 0.2). In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles, peaks of about 7.5, 14.5, 15.1, 20.1, 21.5, and 24.1 (± about 0.2). In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From the 2θ angles are peaks of about 7.5, 15.1, 18.1, 20.1, 20.5, 22.6, and 24.1 (± about 0.2). In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected The peaks are about 7.5, 20.1, and 24.1 (± about 0.2) from the 2θ angle. In some embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected Since having the d value as provided in Table 25 ( The peak. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 6.1, 5.9, 5.1, 4.9, 4.8, 4.6, 4.4, 4.3, 4.1, 4.1, 3.7, 3.6, and 3.0. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 6.1, 5.9, 4.4, 4.1, and 3.7. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) are peaks of about 11.8, 5.9, 4.9, 4.4, 4.3, 3.9, and 3.7. In other embodiments, the XRPD pattern of the crystalline tosylate salt comprises one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks), the peaks selected From d value ) is the peak of about 11.8, 4.4 and 3.7.

In some or any embodiments, the crystalline tosylate exhibits a ¹³ C NMR spectrum substantially corresponding to the spectrum in Figure 12 or exhibits a spectrum having a peak substantially corresponding to Table 28. In some or any embodiments, the crystalline tosylate exhibits a ¹³ C NMR spectrum having one or more peaks (eg, at least 3, at least 4, at least 5, at least 6, or at least 7 peaks) The peaks are selected from the group consisting of about 166.9, 164.3, 162.2, 160.6, 151.8, 149.4, 143.2, 140.2, 139.1, 136.0, 131.8, 129.4, 128.6, 127.7, 123.9, 116.8, 115.1, 112.2, 105.2, 100.3, 58.5, 45.3. Peaks at 37.4 and 23.9 (approximately ±0.2 ppm). In some or any embodiments, the crystalline tosylate salt exhibits at about 151.8, 149.4, 143.2, 136.0, 131.8, 123.9, 116.8, 115.1, 112.2, 105.2, 100.3, 58.5, 45.3, 37.4, and 23.9 (about ± ¹³ C NMR spectrum of the peak at 0.2 ppm). In some or any of the embodiments, crystalline tosylate salt ¹³ exhibits a peak (about ± 0.2 ppm) and at the at about 23.9 to 143.2,136.0,131.8,123.9,112.2,105.2,100.3,58.5,45.3,37.4 C NMR spectrum. In some or any of the embodiments, crystalline tosylate salt exhibit ¹³ C NMR spectrum having a peak (about ± 0.2 ppm) of at about 143.2,136.0,131.8,123.9,112.2,105.2 and 100.3.

In some or any embodiments, the crystalline salt has an XRPD peak at about 2, 4, 15.1, 17.4, 17.8, 18.1, 18.5, 19.1, 20.1, 20.4, 21.5, 22.6, 24.0, 24.8, and 27.0. In some or any embodiments, the crystalline salt has an XRPD peak at about 2, 14.1, 18.1, 19.1, 20.1, 20.4, 21.5, 22.6, and 24.0 at 2 theta angles. In some or any embodiments, the crystalline salt has an XRPD peak at about 2, 11.1, 20.1, 20.4, 22.6, 24.0, and 24.8 at 2 theta angles. In some or any embodiments, the crystalline salt has an XRPD peak at about 2,4, 15.1, 18.1, 20.1, 20.4, 22.6, and 24.0 at 2 theta angle.

In some or any embodiments, the crystalline salt has a d value ( ) is an XRPD pattern of peaks of about 11.9, 5.9, 5.1, 5.0, 4.9, 4.8, 4.6, 4.4, 4.3, 4.1, 3.9, 3.7, 3.6, and 3.3. In some or any embodiments, the crystalline salt has a d value ( ) is an XRPD pattern of peaks of about 11.9, 5.9, 4.9, 4.6, 4.4, 4.3, 4.1, 3.9, and 3.7. In some or any embodiments, the crystalline salt has a d value ( ) is an XRPD pattern of peaks of about 11.9, 5.9, 4.4, 4.3, 3.9, 3.7, and 3.6. In some or any embodiments, the crystalline salt is at a d value ( ) has XRPD peaks at about 11.9, 5.9, 4.9, 4.4, 4.3, 3.9, and 3.7.

In some or any embodiments, crystalline (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5 -yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketotoluenesulfonate exhibits a crystalline polymorph of at least one of the following: solid state ¹³ C NMR spectra at 143.2, 136.0, 131.8, 123.9, 112.2, 105.2, and 100.3 ppm (±) 0.2 ppm) with a peak; an X-ray diffraction pattern comprising the following d values ( Characteristic peaks indicated: 11.9, 5.9, 4.9, 4.4, 4.3, 3.9, and 3.7; and X-ray diffraction patterns including 2θ angles of 7.4, 15.1, 18.1, 20.1, 20.4, 22.6, and 24.0 (±0.2°) Peak reflection at 2θ).

In some embodiments, the crystalline tosylate exhibits a single endothermic peak between room temperature and about 350 ° C in differential scanning calorimetry, wherein the single endothermic peak maximum occurs from about 320 ° C to about 335 ° C. between. In certain embodiments, a single endothermic peak maximum occurs between about 330 °C and about 335 °C. In some embodiments, a single endothermic peak maximum occurs from about 333 °C to about 334 °C.

In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The crystalline tosylate salt of -3( 7H )-ketone has no observable endotherm from about 25 ° C to about 250 ° C as determined by DSC. In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The DSC thermogram of the crystalline tosylate salt of -3( 7H )-ketone comprises a maximum between about 320 ° C and about 335 ° C, between 330 ° C and about 335 ° C or between about 333 An endotherm between °C and about 334 °C. In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The DSC thermogram of the crystalline tosylate salt of -3( 7H )-ketone substantially corresponds to the DSC thermogram of Figures 2a, 3a, 4a, 5a or 7a. In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The DVS isotherm of the crystalline tosylate salt of -3( 7H )-ketone substantially corresponds to the DVS isotherm plot of Figure 9. In certain embodiments, (8 S ,9 R) -5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The crystalline tosylate salt of -3( 7H )-ketone does not exhibit significant weight change (eg, less than about 0.05 wt%, less than about 0.1 wt%, less than about 0.15 wt) at about 0% to about 95% relative humidity. % or less than about 0.2 wt%).

In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The TGA thermogram of the crystalline tosylate salt of -3( 7H )-ketone substantially corresponds to the TGA thermogram of Figures 2b, 3b, 4b, 5b or 7b. In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The TGA thermogram of the crystalline tosylate salt of -3( 7H )-ketone is heated from about room temperature to about 200 ° C, about 210 ° C, about 220 ° C, about 230 ° C, about 240 ° C, about 250 ° C. Not exhibiting significant weight loss (eg, less than about 0.05 wt%, at temperatures of about 260 ° C, about 270 ° C, about 280 ° C, about 290 ° C, about 300 ° C, about 310 ° C, about 320 ° C, or greater than about 320 ° C, Less than about 0.1 wt%, less than about 0.5 wt%, less than about 1 wt%, less than about 5 wt%, less than about 10 wt%, less than about 15 wt%, less than about 20 wt%, or less than about 25 wt%. In various embodiments, the crystalline tosylate salt provided herein has a weight loss between about 25 ° C and about 200 ° C in the thermogravimetric analysis chart of no greater than about 1%, no greater than about 0.5%, or no greater than About 0.1%.

In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The crystalline tosylate salt of -3( 7H )-one was not solvated. In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The crystalline tosylate salt of -3( 7H )-one is anhydrous. In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The crystalline tosylate salt of -3( 7H )-ketone is thermally stable. In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The crystalline tosylate salt of -3( 7H )-ketone is non-hygroscopic.

In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4, as provided herein -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The crystalline tosylate salt of -3( 7H )-ketone comprises, the preparation, treatment and/or storage of the ( 8S , 9R )-5-fluoro-8-(4-fluorophenyl)-9-( 1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The pharmaceutical composition or pharmaceutical product of the -3( 7H )-ketone crystalline tosylate exhibits desirable properties.

In another aspect, the synthesis of (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5- ,8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketone method. Scheme A outlines an example of a synthetic method. In each of its embodiments, the method comprises three steps, the first step of which is 5-fluoro-2-(2-(1-methyl-1H-1,2,4-triazol-5-yl) Ethyl benzyl 3-nitrobenzoate (ie,

Contacting 4-fluorobenzaldehyde in a mixture comprising one or more of the steps (a) solvent and titanium (III) chloride to produce a first intermediate (b in Scheme A). In certain embodiments, the one or more steps (a) solvent is selected from the group consisting of THF and MeOH, for example, a volume/volume ratio of 6 parts THF: 1 part MeOH. Titanium (III) chloride can be added to the first reaction mixture at a temperature of from 0 °C to room temperature.

Option A

In some embodiments, the agitation comprises 5-fluoro-2-(2-(1-methyl-1 H -1,2,4-triazol-5-yl) at a temperature of from about 30 ° C to about 50 ° C. a reaction mixture of methyl hydrazide)-3-nitrobenzoate, one or more steps (a) a solvent and titanium (III) chloride.

The second step of the synthetic method provided (step (b)) separates the enantiomer (c) of the first intermediate (b) by separation of the palms. The splitting of the first intermediate (b) into its enantiomers can be carried out by any method known to those skilled in the art, such as, for example, high performance liquid chromatography and supercritical fluid chromatography. Equal chromatography method. The enantiomers shown as c in Scheme A can be separated in the second step of the synthesis process.

The third step of the synthetic method (step (c)) comprises contacting the separated enantiomer (c) of the first intermediate (b) with one or more steps (c) of a solvent and hydrazine monohydrate to produce (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9 -dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketone. In certain embodiments, the one or more steps (c) are independently selected from the group consisting of methanol, ethanol, and acetonitrile. In certain embodiments, the solvent of step (c) is methanol. In certain embodiments, the solvent of step (c) is ethanol. In other embodiments, the solvent of step (c) is acetonitrile. Typically, the third step can be carried out at room temperature with the reactants allowed to stand overnight.

Alternatively, the intermediate b is treated with a solvent such as methanol, ethanol or acetonitrile and hydrazine monohydrate to give 5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2 ,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketone, which is then resolved into (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-A by palmar splitting Base-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketone. The palm-shaped splitting can be carried out by any method known to those skilled in the art, for example, chromatographic methods such as high performance liquid chromatography, supercritical fluid chromatography, and simulated moving bed chromatography.

In another aspect, the preparation of (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5- ,8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketotosylate method. The process for preparing tosylate salts provided herein is suitable for large scale production of tosylate salts and can meet GMP requirements.

In certain embodiments, the method of preparation comprises (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1) in one or more solvents H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketone free base is contacted with p-toluenesulfonic acid and one or more solvents are removed to produce ( 8S , 9R )-5-fluoro-8-(4-fluorophenyl)- 9-(1-Methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate. In certain embodiments, the free base is used in the synthesis of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H as described herein. -1,2,4-triazol-5-yl)-8,9-dihydro- 2H -pyrido[4,3,2-de]呔 -3( 7H )-ketone method to obtain.

In some or any of the embodiments of the method of preparing the tosylate salt form, the free base is suspended or dissolved in one or more solvents independently selected from the group consisting of THF, acetone, methanol, acetonitrile, and DCM. In certain embodiments of the process for the preparation of the tosylate form, the free base is suspended or dissolved in a mixture of methanol and acetonitrile. In certain embodiments of the process for the preparation of the tosylate form, the free base is suspended or dissolved in a mixture of DCM and acetonitrile. In certain embodiments of the process for the preparation of the tosylate form, the free base is suspended or dissolved in a mixture of acetone and THF. In certain embodiments of the process for the preparation of the tosylate form, the free base is suspended or dissolved in acetone. In certain embodiments of the process for the preparation of the tosylate form, the free base is suspended or dissolved in THF. In certain embodiments of the process for the preparation of the tosylate form, the free base is suspended or dissolved in one or more solvents independently selected from the group consisting of THF, acetone, methanol, acetonitrile, and DCM, and heated prior to the addition of TsOH. In certain embodiments of the process for the preparation of the tosylate form, the free base is suspended or dissolved in a mixture of acetone and THF and heated prior to the addition of TsOH. In some embodiments, the free base is suspended in a solvent consisting essentially of THF. In certain embodiments of the process for the preparation of the tosylate form, the free base is suspended or dissolved in THF and heated prior to the addition of TsOH.

In some or any embodiments, the p-toluenesulfonic acid is dissolved or suspended in one or more solvents and then added to the free base suspended or dissolved in the second set of one or more solvents. In some or any embodiments, the free base is dissolved or suspended in one or more solvents and then added to p-toluenesulfonic acid suspended or dissolved in a second group of one or more solvents.

In some or any embodiments, (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4- is raised at elevated temperature Triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 The -3(7H)-one is contacted with p-toluenesulfonic acid. In some or any embodiments, the TsOH and the free base can be contacted at a temperature between about 0 °C and about 70 °C. Typically, the temperature of the free base upon contact with TsOH is from about 20 ° C to 55 ° C. In some or any embodiments, the free base is contacted with TsOH at a temperature of from about 30 ° C to about 70 ° C, from about 25 ° C to about 30 ° C, from about 30 ° C to about 40 ° C, from about 40 ° C to about 50 ° C. From about 50 ° C to about 60 ° C, from about 60 ° C to about 70 ° C or from about 48 ° C to about 58 ° C. The solvent can be removed and the salt dried according to methods known to those skilled in the art.

In some or any embodiments, after contacting the free base with TsOH, the resulting solution/suspension is allowed to stand under conditions sufficient to precipitate the crystalline form and the crystalline form is isolated. In some or any embodiments, the conditions sufficient to precipitate the crystalline form comprise cooling. In some or any embodiments, the conditions sufficient to precipitate the crystalline form comprise cooling to a temperature of 25 ° C or less.

In addition to the methods provided herein, the (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl) provided herein can also be prepared using techniques known in the art. -1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 a solid form of -3( 7H )-ketone, including but not limited to, melt cooling, rapid melt cooling, freeze drying, spray drying, drum drying, lyophilization, melt quenching, solvent rapid Evaporation, slow solvent evaporation, solvent recrystallization, slurry recrystallization, melt crystallization, desolvation, sublimation, recrystallization in confined spaces (eg, in nanopores or capillaries), on surfaces or stencils (eg Recrystallization on the polymer, recrystallization in the presence of additives (eg, eutectic counter molecules), dehydration, rapid cooling, slow cooling, vapor diffusion, grinding, freeze milling, solvent micromilling, microwave induced precipitation, ultrasonication Treatment induced precipitation, laser induced precipitation and precipitation from supercritical fluids.

Another aspect is the preparation of (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)- 8,9-Dihydro-2H-pyrido[4,3,2-de]呔 a method of -3(7H)-ketotosylate, which comprises

Step (1): (8 S , 9 R )-5-fluoro-8 in the presence of one or more solvents selected from the group consisting of THF, acetone, methanol, acetonitrile and DCM at elevated temperature -(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3 ,2-de]呔 -3(7 H )-one is contacted with p-toluenesulfonic acid;

Step (2): allowing it to stand under conditions sufficient to precipitate the crystalline form;

Step (3): Separation of the crystalline form.

In some or any embodiments, the elevated temperature is from about 30 °C to about 70 °C. In some or any embodiments, the one or more step 1 solvents are independently selected from the group consisting of methanol and acetonitrile. In some or any embodiments, the one or more Step 1 solvents are independently selected from the group consisting of DCM and acetonitrile. In some or any embodiments, the one or more step 1 solvents are independently selected from the group consisting of acetone and THF. In some or any embodiments, the solvent of step 1 is acetone. In some or any embodiments, the solvent of step 1 is THF. In some or any embodiments, the conditions sufficient to precipitate the crystalline form comprise cooling. In some or any embodiments, the conditions sufficient to precipitate the crystalline form comprise cooling to a temperature of 25 ° C or less. In some or any embodiments, the method further includes

Step (a): make

Contacting 4-fluorobenzaldehyde in a mixture comprising one or more steps (a) a solvent and titanium (III) chloride to prepare a first intermediate;

Step (b): separating the enantiomer of the first intermediate by separating the palms; and

Step (c): contacting the separated enantiomer of the first intermediate with the monohydrate hydrazine in one or more of the solvents of step (c) to produce (8 S , 9 R )-5-fluoro-8- (4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3, 2-de]呔 -3(7 H )-ketone. In some or any embodiments, the separated enantiomers of the first intermediate are contacted with the monohydrate hydrazine in one or more solvents (c) independently selected from the group consisting of methanol, ethanol, and acetonitrile. In some or more embodiments, the one or more steps (a) are independently selected from the group consisting of THF and methanol. In some or any embodiments, the method further includes

Step (x): make

Contact with monohydrate in one or more step x solvents to produce 5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazole-5- ,8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-one; and step (y): separating (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl- by palm separation 1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-one. In some or any embodiments, the one or more steps (x) are independently selected from the group consisting of methanol, ethanol, and acetonitrile.

In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9, prepared according to the methods set forth in the preceding paragraphs or as substantially set forth in the Examples below, is provided. -(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate. For example, in some embodiments, (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-) is provided which is crystallized from a solvent consisting essentially of tetrahydrofuran (THF). 1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-ketotosylate.

In some or any embodiments, the tosylate exhibits a thermal weight loss of less than 2% by TGA at a temperature of about 280 ° C or less.

In some or any embodiments, the tosylate salt exhibits a hysteresis value of less than about 1% at 25 ° C and RH0% to RH 95% in DVS.

Composition comprising a pharmaceutical composition

In yet another aspect, provided is substantially pure (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2, 4-triazol-5-yl)-8,9-dihydro- 2H -pyrido[4,3,2-de]呔 a composition of or consisting essentially of -3( 7H )-ketotosylate (in its other embodiment, in its crystalline form as provided herein).

In some embodiments, provided is (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole- 5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 A pharmaceutical composition of -3( 7H )-ketotosylate (containing a crystalline form thereof as described herein) and a pharmaceutically acceptable excipient and/or carrier. The choice of excipient will depend to a large extent on, for example, the following factors: the particular mode of administration, the effect of the excipient on the solubility and stability of the active ingredient, and the nature of the dosage form.

The pharmaceutical compositions provided herein can be provided in unit dosage form or in multiple dosage forms. As used herein, unit dosage form refers to physically discrete units as are known in the art to be suitable for administration to humans and animal subjects and for individual packaging. Each unit dose contains a predetermined amount of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2 sufficient to produce the desired therapeutic effect. ,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketotosylate and the desired pharmaceutical carrier or excipient. Examples of unit dosage forms include ampoules, cartridges, and individual packaged lozenges and capsules. The unit dosage form can be administered in portions or multiple times. Multiple dosage forms are a plurality of identical unit dosage forms that are packaged in a single container and administered in separate unit dosage forms. Examples of multiple dosage forms include vials, vials or capsules, or pint bottles or gallons.

(8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9 -dihydro-2 H -pyrido[4,3,2-de]呔 The amount of -3( 7H )-ketotosylate in unit dosage form can be, for example, from about 5 μg to about 1500 μg, from about 20 μg to about 1250 μg, from about 25 μg to about 1000 μg or From about 25 μg to about 250 μg. In some embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5- ,8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The amount of -3( 7H )-ketotosylate in a unit dosage form comprises (wherein the following does not include the mass of the tosylate moiety contribution in the salt) from about 5 μg to about 30 μg, from about 20 μg to about 60 μg. From about 50 μg to about 100 μg, from about 120 μg to about 250 μg, from about 20 μg to about 112 μg or from about 25 μg to about 250 μg of (8 S , 9 R )-5-fluoro-8 -(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3 ,2-de]呔 -3(7 H )-ketone. In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The amount of -3( 7H )-ketone is about 10 μg, about 20 μg, about 25 μg, about 30 μg, about 50 μg, about 75 μg, about 100 μg, about 150 μg, about 200 μg, about 250 μg. Or about 1000 μg.

Oral dosage form. In one embodiment, a pharmaceutical composition as provided herein is formulated for oral administration to an individual. Pharmaceutical compositions suitable for oral administration can be presented in discrete dosage forms such as, but not limited to, lozenges, chewable lozenges, caplets, capsules and liquids (for example, flavored syrups). These dosage forms contain a predetermined amount of active ingredient and may be prepared by methods of pharmacy known to those skilled in the art. Generally, see Remington's Pharmaceutical Sciences , 18th ed., Mack Publishing, Easton PA (1990).

For example, the oral dosage forms provided herein can be prepared by combining (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-A according to conventional pharmaceutical mixing techniques. Base-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 ( 7H )-ketotosylate (in one embodiment, in its crystalline form as provided herein, hereinafter "active ingredient") is in admixture with at least one excipient. The excipients can take a wide variety of forms depending on the form of preparation desired to be administered. Because tablets and capsules are easy to administer, they represent the most advantageous oral dosage unit form, in which case solid excipients are employed. If desired, the tablet can be coated by standard aqueous or non-aqueous techniques. These dosage forms can be prepared by any of the methods of pharmacy. In general, pharmaceutical compositions and dosage forms are prepared by uniformly and intimately admixing the active ingredient with a liquid carrier, a fine solid carrier or both, and then, if necessary, shaping the product into the desired presented form. A disintegrant or lubricant can be used in the pharmaceutical compositions and dosage forms of the invention. The production of the pharmaceutical composition or dosage form of the present invention may require, in addition to the therapeutic pharmaceutical ingredient, excipients or additives, including but not limited to diluents, binders, lubricants, disintegrants, colorants, flavoring agents, Sweeteners and the like or mixtures thereof. By incorporating these and other additives, various dosage forms (e.g., tablets, capsules, caplets, tablets, and the like) can be prepared. Such dosage forms comprise, for example, hard gelatin capsules, caplets, dragees, enteric coated lozenges (for example, for delayed action), multiple compressed lozenges, long acting lozenges, solution lozenges, effervescent ingots Agents, buccal and sublingual tablets, tablets and the like. Dosage forms or dosage formulations can be formed by methods well known in the art. See, for example, Remington's Pharmaceutical Sciences , 16th and 18th editions, Mack Publishing Company, Easton, Pa. (1980 and 1990). See also US Pharmacopeia XXI , US Pharmacopeia Convention, Rockville, Md. (1985).

Other Dosage Forms. In certain embodiments, a pharmaceutical composition as provided herein can be formulated in a parenteral dosage form. Parenteral dosage forms can be administered to a patient by a variety of routes including, but not limited to, subcutaneous, intravenous (including bolus), intramuscular, and intraarterial. Because such administration typically bypasses the patient's natural defense against contaminants, the parenteral dosage form is sterile in some embodiments or can be sterilized prior to administration to the patient. Examples of parenteral dosage forms include, but are not limited to, injectable solutions, dissolved or suspended in pharmaceutically acceptable vehicles, i.e., injectable dry products, i.e., injectable suspensions and emulsions. Suitable vehicles which can be used to provide the parenteral dosage forms provided herein are well known to those skilled in the art.

In yet another embodiment, a pharmaceutical composition as provided herein can be formulated in a transdermal, topical or mucosal dosage form. The percutaneous, topical, and mucosal dosage forms provided herein include, but are not limited to, ophthalmic solutions, sprays, aerosols, creams, lotions, ointments, gels, solutions, emulsions, suspensions, or those skilled in the art. Other forms are known. See, for example, Remington's Pharmaceutical Sciences , 16th and 18th edition, Mack Publishing, Easton PA (1980 &1990); and Introduction to Pharmaceutical Dosage Forms , 4th edition, Lea & Febiger, Philadelphia (1985). Suitable excipients (e.g., carriers and diluents) and other materials that can be used to provide the transdermal, topical, and mucosal dosage forms encompassed by the present disclosure are well known to those skilled in the art and are intended to be given Depending on the particular organization of the pharmaceutical composition or dosage form.

treatment method

Provided herein are methods of treating a disease or condition or a symptom thereof, as explained below.

In some or any embodiments, provided herein is (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-tri Zyrid-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 Use of a -3( 7H )-ketotoluenesulfonate for treating a disease or condition of an individual or a symptom thereof.

In some or any embodiments, the method of treating cancer or a symptom thereof comprises administering to a subject having cancer a therapeutically effective amount of ( 8S , 9R )-5-fluoro-8-(4-fluorophenyl)-9 -(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate.

Some embodiments or any of the embodiments provide a method of enhancing cytotoxic cancer therapy in an individual in need of treatment comprising administering to the subject a therapeutically acceptable amount of (8 S , 9 R )-5-fluoro-8 -(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3 ,2-de]呔 -3(7 H )-ketotosylate.

In some or any embodiments, provided herein are methods for treating cancer comprising administering to a subject in need of treatment a therapeutically effective amount of ( 8S , 9R )-5-fluoro-8-(4-fluorophenyl) 9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketotosylate and ionizing radiation and/or one or more chemotherapeutic agents. In some or any embodiments, the compounds described herein are administered simultaneously with ionizing radiation and/or one or more chemotherapeutic agents. In some or any embodiments, the compounds described herein are administered sequentially with ionizing radiation and/or one or more chemotherapeutic agents. Ionizing radiation and chemotherapeutic agents are known to those skilled in the art.

In some or any embodiments, the therapeutic agent is an alkylating agent, such as methyl methanesulfonate (MMS), temozolomide, and dacarbaZine (DTIC); topoisomerase 1 inhibitors, for example Topotecan, Irinotecan, Rubitecan, Exatecan, Lurtotecan, Gimetecan, Diflomotecan (homocamptothecin), 7-substituted non-silatecan, 7-carboxycylcamptothecin (BNP 1350), SN38, NK012 (by making SN-38 covalent SN-38-release nanodevices attached to the block copolymer PEG-PG1u and then self-assembled amphiphilic block copolymer in an aqueous medium) and XR 11576/MLN 576; alemtuzumab (alemtuzumab) Arsenic trioxide; aspartame (pegylated or non-PEGylated); bevacizumab; cetuximab; platinum-based compounds such as cisplatin , carboplatin, oxaliplatin and triplatinum tetrachloride; cladribine; daunorubicin; Adriamycin (doxorubicin); Ida adriamycin (idarubicin); fludarabine (fludarabine); 5- fluorouracil; gemtuzumab (gemtuzumab); methotrexate (methotrexate); Paclitaxel ^TM; paclitaxel (the Taxol) Dimensions; thioguanine; hormone therapy, such as antiestrogens, antiandrogens and gonadotropin-releasing hormone analogues; interferons, such as alpha interferon; nitrogen mustard, such as bucacaine sulfate (busulfan) ), melphalan and mechlorethamine; retinoids such as tretinoin; tyrosine kinase inhibitors such as gefitinib and kilikeril Imatinib); a proteasome inhibitor, such as bortezomib; or an agent for treating the signs or symptoms induced by this therapy, including allopurinol, filgrastim, granisetron/angon Ondansetron / palonosetron and dronabinol.

In some or any embodiments, a method for treating cancer comprising administering to a subject in need of treatment a therapeutically effective amount of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl) -9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketotosylate and topoisomerase inhibitor. In some or any embodiments, the topoisomerase inhibitor is SN38, anti-cancer or NK012. In some or any embodiments, the topoisomerase inhibitor is anti-cancer. In some or any embodiments, the cancer is breast cancer.

In some or any embodiments, a method for treating cancer comprising administering to a subject in need of treatment a therapeutically effective amount of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl) -9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate and platinum. In some or any embodiments, the platinum is cisplatin, carboplatin, yuleplatin or triplatinum tetranitrate. In some or any embodiments, the topoisomerase inhibitor is cisplatin. In some or any embodiments, the cancer is breast cancer.

In some or any embodiments, a method of treating cancer comprising administering to a subject in need of treatment a therapeutically effective amount of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9 -(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate and Dimensions. In some or any embodiments, the cancer is colorectal cancer.

In some embodiments, provided herein are methods of treating cancers that are defective in homologous recombination (HR)-dependent DNA double-strand break (DSB) repair pathways, comprising administering to a subject in need of treatment a therapeutically effective amount of (8 S , 9 R -5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate. In certain embodiments, the cancer comprises one or more cancer cells that are reduced or eliminated relative to normal cells by the ability of HR to repair DNA DSB. In some embodiments, the cancer cell has a BRCA1 or BRCA2 deficient phenotype. In some embodiments, the cancer cells are defective in BRCA1 or BRCA2. In some embodiments, the methods provided herein involve treating an individual mutated to a heterozygous gene encoding a component of a HR dependent DNA DSB repair pathway. In one embodiment, the individual is heterozygous for the mutation in BRCA1 and/or BRCA2.

In certain embodiments, the cancer to be treated includes homologous phosphatase and tensin (PTEN) deficient cells (eg, PTEN mutations or cells that exhibit little or no presence). In certain embodiments, the cancer to be treated comprises a cell having a mutation in the PTEN gene. Exemplary cancers associated with PTEN deficiency can include, for example, glioblastoma, endometrial cancer, prostate cancer, lung cancer, and breast cancer.

In certain embodiments, the cancer to be treated is caused by an activating mutation in the Wnt signaling pathway ("Wnt-mediated cancer"). It should be understood that "activating mutation of the Wnt signal transduction pathway" is intended to include, for example, an oncogene mutation in the gene that causes β-catenin to accumulate in cancer cells, and the CTNNB1 gene (which encodes β-catenin). A gain-of-function mutation, a mutation in an APC tumor suppressor gene, or a mutation in the AXIN2 gene. For example, it is believed and is not intended to be limited by any theory or mechanism, and the treatment available for Wnt-mediated cancer can be achieved by inhibiting tankyrase (an enzyme with poly(ADP-ribose) polymerase activity). . Exemplary Wnt-mediated cancers that can be treated according to the methods provided herein include, for example, bladder cancer, breast fibromatosis, cervical cancer, colorectal cancer, colon cancer, dural fibroma, esophageal adenocarcinoma, familial adenoma Polyposis, fundus glandular polyps, gastric cancer, gastric adenoma, gastrointestinal carcinoid tumor, hepatic blastoma, hepatocellular carcinoma, adolescent nasopharyngeal angiofibroma, non-Hodgkin's lymphoma, lung adenocarcinoma, neural tube blast Tumor, melanoma, ovarian cancer, pancreatic cancer (including, for example, non-catheter solid pseudopapillary tumors and non-catheter adenocarcinoma), pancreatic blastoma, hairy stromal tumor, prostate cancer, small intestine adenocarcinoma, synovial sarcoma, Thyroid cancer, cervical cancer, endometrial cancer, and Wilm's tumor.

In some embodiments of the methods of treating cancer or a symptom thereof provided herein, cancer is bladder cancer, breast cancer (including metastatic breast cancer, BRCA positive breast cancer, and BRCA negative breast cancer), cervical cancer, colon cancer, colorectal cancer, EBV-related Tumors (including Burkitt's lymphoma, nasopharyngeal carcinoma, lymphoma in AIDS patients, smooth muscle tumors in AIDS patients, Hodgkin's disease, non-Hodgkin's lymphoma, lymphoid hyperplasia in immunosuppressed patients) Lymphoma, EBV+gastric cancer, EBV+ breast cancer, T-cell lymphoma, endometrial cancer (including cancer and sarcoma), gastrointestinal stromal tumor, glioma, glioblastoma ) pleomorphic glioblastoma and degenerative astrocytoma), head and neck cancer, hepatocellular carcinoma, hereditary nonpolyposis colon cancer (HNPCC), kidney cancer (kidney cancer), leukemia (including acute myeloid) Leukemia, chronic lymphocytic leukemia), lung cancer (including non-small cell lung cancer, small cell lung cancer), lymphoma (including mantle cell lymphoma), cholangioblastoma, melanoma, meningococcal tumor, Myelodysplastic syndrome, ovarian cancer (including advanced ovarian cancer, highly differentiated serous ovarian cancer, platinum-sensitive ovarian cancer, platinum-resistant ovarian cancer, platinum-refractory ovarian cancer, and BRCA-negative ovarian cancer), pancreatic cancer (including BRCA negative) Pancreatic cancer), peritoneal cancer, prostate cancer (including BRCA-negative prostate cancer, metastatic prostate cancer, and castration-resistant prostate cancer), renal cancer, thyroid cancer, uterine cancer sarcoma, or uterine cancer.

In certain embodiments, provided herein are methods of treating a disease or condition associated with a PTEN deficiency or a symptom thereof, comprising administering to a subject having a disease or condition associated with a PTEN deficiency a therapeutically effective amount (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate. Exemplary diseases and conditions associated with PTEN deficiency include, for example, Cowden syndrome, Bannayan-Riley-Ruvalcaba syndrome, and Lylmite-Dukelo Disease (Lhermitte-Duclos disease), Proteus syndrome, Protos-like syndrome or hamartoma syndrome.

Certain embodiments provide methods of treating an inflammatory disease in an individual in need of treatment, including but not limited to arthritis, gout, inflammatory bowel disease, CNS inflammation, multiple sclerosis, allergic brain Inflammation, sepsis, septic shock, hemorrhagic shock, pulmonary fibrosis, and uveitis, the method comprising administering to the individual a therapeutically acceptable amount of (8 S , 9 R )-5-fluoro-8- (4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3, 2-de]呔 -3(7 H )-ketotosylate.

Certain embodiments provide methods of treating an immune disease or condition, such as rheumatoid arthritis and septic shock, in an individual in need of treatment, comprising administering to the individual a therapeutically acceptable amount (8 S , 9 R -5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate.

Certain embodiments provide methods of treating a degenerative disease in an individual in need of treatment, including but not limited to diabetes and Parkinson's disease, the method comprising administering to the individual a therapeutically acceptable (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)- 8,9-Dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate.

In certain embodiments of this use, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2, 4-triazol-5-yl)-8,9-dihydro- 2H -pyrido[4,3,2-de]呔 The exact amount of -3( 7H )-ketotosylate depends on the patient's state of health, body weight and the like. In some embodiments, it is believed that such therapeutically effective amounts should be determined by a caregiver by routine experimentation (eg, a dose escalation clinical trial). In certain embodiments, when used in a patient, the effective amount for such use will depend on the severity and course of the disease, condition or condition, prior therapy, the health of the patient and the drug's response, and the judgment of the treating physician. .

In certain embodiments of the methods of treatment provided herein, including those provided above, (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9 can be administered in solid form -(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-one tosylate. In some embodiments, the salt is administered in a crystalline form.

In some embodiments of the methods of treatment provided herein, the individual is administered (8 S in a regimen of about 1 week, about 2 weeks, about 3 weeks, about 1 month, about 2 months, or about 6 months). ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro -2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate. In some embodiments, the tosylate salt is administered daily as per the protocol. In other embodiments, the tosylate salt is administered 2 days, 3 days, or 4 days per week, during which time the tosylate salt is not administered on a weekly basis for several days.

In some cases, administration of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole -5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketotosylate (including administration in the form of a crystalline form), the condition of the patient is not improved or significantly improved, and depending on the condition, long-term (ie, extended time, included) The compound is administered throughout the life of the patient to improve or otherwise control or limit the symptoms of the disease or condition of the patient.

In some cases, if the condition of the patient has not improved or has not been significantly improved, the active ingredient will continue to be administered as appropriate according to the doctor's advice; or, depending on the circumstances, the dose of the administered drug will be temporarily reduced or temporarily suspended for a period of time ( That is, "drug holiday"). In certain embodiments, the length of the drug holiday varies between 2 days and 1 year, including (for example only) 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 Days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days or 365 days.

In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The amount of -3( 7H )-ketotoluenesulfonate corresponding to an effective amount varies depending on factors such as the specific compound, the disease or condition and its severity, the personality of the individual or subject in need of treatment (eg, body weight). . However, in some embodiments, an effective amount is determined based on the particular circumstances of the case, including, for example, the particular agent being administered, the route of administration, the condition being treated, and the individual or subject being treated. However, in certain embodiments, the dosage for adult treatment ranges from about 5 μg to about 8000 μg per day, and in a particular embodiment from about 10 μg to about 5000 μg per day. In certain embodiments, (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-tri Zyrid-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The amount of -3( 7H )-ketotosylate is between about 50 μg to about 5000 μg, between about 50 μg to about 1500 μg, between about 50 μg to about 1000 μg, Between about 50 μg to about 500 μg, between about 50 μg to about 250 μg, between about 50 μg to about 200 μg, between about 50 μg to about 150 μg, between Between about 50 μg to about 100 μg, between about 25 μg to about 2500 μg, between about 25 μg to about 1000 μg, between about 25 μg to about 250 μg, between about 25 μg Between μg and about 150 μg or between about 25 μg to about 75 μg. In various embodiments, the desired dose may conveniently be administered in a single dose or simultaneously (or in a short period of time) or at appropriate intervals (eg, two, three, four or more divided doses per day). To present.

In certain embodiments, (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-tri Zyrid-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 A suitable daily dose of -3( 7H )-ketotosylate is from about 0.1 μg/kg body weight to about 200 μg/kg body weight. In certain embodiments, the daily dose is from about 0.3 μg/kg body weight to about 3.0 μg/kg body weight. In some embodiments, the specified daily dose in a larger individual (including, but not limited to, a human) is in the range of from about 25 μg to about 8000 μg, conveniently in divided doses (including, but not limited to, up to four times per day) ) or in the form of extended release. In certain embodiments, suitable unit dosage forms for oral administration include the above dosage forms. Since the number of variables for individual treatment regimens is large, the above ranges are merely illustrative and often significantly shift the suggested values. In certain embodiments, the dosage end varies depending on a number of variables that are not limited to the activity of the compound used, the disease or condition being treated, the mode of administration, the needs of the individual individual, the severity of the disease or condition being treated. Sex and the judgment of the practitioner.

Instance

Methods and Procedures. The reagents and solvents used below are available from commercial sources such as Aldrich Chemical Company (Milwaukee, Wis., USA). Routine chemical analysis was carried out using NMR, MS and HPLC. The effective NMR peaks are tabulated by chemical shift and according to multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br s, broad singlet) and number of protons To mark. Mass spectral data on the mass M of the parent ion is provided. HPLC data is provided as a percentage of purity.

XRPD patterns were obtained on a Bruker D8 Advance diffractometer (Bruker AXS, Madison, WI, USA) unless otherwise stated. Gently tile the sample onto the zero background 矽 insert sample holder. Continuous 2θ scan from 4° to 40° using Cu Kα (λ 1.54056) Radiation source and generator power of 40 kV and 40 mA. A 2θ step of 0.05 degrees/step and a step time of 1 second/step are used. The experiment was carried out at room temperature and ambient humidity. The standard error is about 0.2 2θ angle. The entire list of peaks identified in the XRPD pattern or a subset thereof may be sufficient to characterize the polymorph obtained.

A TA Instruments Q2000 Differential Scanning Calorimeter (New Castle, DE, USA) was used to obtain the DSC thermograms unless otherwise stated. The sample was weighed directly into a DSC aluminum pan. The disk is sealed by applying pressure by hand and pushing each part of the disk together (also known as a loose cover configuration). The temperature ramped from 25.00 °C to 400.00 °C at 10.00 °C/min, except as explained below. In the DSC diagram, an exothermic event is drawn in the upward direction.

TGA thermograms were acquired using a TA Instruments Q500 Thermogravimetric Analyzer (New Castle, DE, USA) unless otherwise stated. The sample was weighed into the pan. The temperature ramped from 25.00 °C to 400.00 °C at 10.00 °C/min, except as explained below.

Unless otherwise stated, DVS was obtained using a standard procedure on DVS ADVANTAGE Type 1 (Alperton, Middlesex, UK). The moisture adsorption desorption isotherm is drawn as outlined below. The standard isotherm runs in a cycle starting at RH 0% from RH 95% at 95% and then drying to RH 0% at 5% RH intervals.

(8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-di Hydrogen-2 H -pyrido[4,3,2- de ]呔 Exemplary synthesis of -3(7 H )-ketone

This example provides the preparation of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4 according to one aspect of the present disclosure. -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 A representative method of -3( 7H )-ketone. The synthesis is outlined in Scheme A.

Preparation of b. A (5 g, 15.5 mmol, 1 eq.) and 4-fluorobenzaldehyde 2 (3.6 g, 29 mmol, 1.87 eq.) in THF (30 mL) and MeOH. Titanium (III) chloride (20% w/w solution, stored in 2 N hydrochloric acid) (80 mL, 6 equivalents) was added dropwise to the suspension in a mixture of (5 mL). The reaction mixture was stirred at 30 to 50 ° C for 2 h. The mixture was then diluted with water (160 mL) and EtOAc (EtOAc) The organic layer was washed merger (100 mL × 3) and washed with sat NaHCO ₃ (50 mL × ₃₎ and _aqueous NaHSO4, dried over Na ₂ SO _4, was concentrated to obtain a yellow solid with petroleum ether (120 mL) of crude solid. The title compound (5.9 g, yield: 95%, purity: 97%). LC-MS (ESI) m / z: 399 (M + 1) +. ¹ H-NMR (400 MHz, CDCl ₃ -d) δ (ppm): 3.58 (s, 3H), 3.87 (s, 3H), 4.16 - 4.19 (d, J ² = 13.2 Hz, 1H), 4.88 (s) , 1H), 5.37-5.40 (d, J ² = 13.2 Hz, 1H), 6.47-6.53 (m, 2H), 6.97-7.01 (m, 2H), 7.37-7.41 (m, 2H), 7.80 (s, 1H).

Compound b was also prepared according to a modified version of the above procedure, wherein the washed combined organic layers were concentrated to 1-2 vol, and then 6 vol heptane was added, after which washing, concentration and addition of heptane were repeated. The solution was then concentrated to 1-2 vol and 6 vol MTBE was added. The mixture was stirred for 1 hour and filtered to give a pale yellow solid (yield: 92% yield, 98% purity).

Preparation of c. Supercritical fluid chromatography (SFC) was used to separate b from the palm to obtain c. A sample was prepared by using methanol as a solvent (b: 45 mg/mL) by heating to 40-50 ° C before injection and filtering. A CHIRALPAK IC, 250 x 30 mm (ID) column and a CO ₂ /methanol (80/20) mobile phase with a flow rate of 65 g/min. The column temperature was maintained at 35 °C. The fraction was expected to flow out of the column as a first peak with a residence time of 2.3 minutes and a residence time of 4.3 minutes for the other enantiomer. UV detection was performed at 254 nm. The recovery of c is about 92% and ee > 98%.

(8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro -2H-pyrido[4,3,2-de]呔 -3(7H)-ketone. To the methanol solution of c was added hydrazine monohydrate, and the mixture was stirred at 25 ° C for 10 hr. The mixture was then filtered and dried to give the compound as a white solid. In some cases, if acetonitrile (10 vol) containing 3 equivalents of hydrazine monohydrate is used instead of methanol, the reaction can be completed in 5 hours at 35 °C. The yield of these procedures is from about 77% to 80%.

(8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl) -8,9-dihydro -2H-pyrido[4,3,2-de]呔 Preparation and comparison of -3(7H)-keto-salt and free base forms

The following example provides (8 S , 9 R )-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8, 9-dihydro-2 H -pyrido[4,3,2- de ]呔 Preparation and characterization of the -3( 7H )-keto salt and free base polymorph. Showing (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8, 9-dihydro-2 H -pyrido[4,3,2- de ]呔 The properties of the -3( 7H )-ketotosylate are superior to those of the other salts and free base forms of the compound. Such improved properties include, but are not limited to, the presence of a single crystal form, no solvation, high melting point, non-hygroscopicity, and/or thermal stability.

(8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro -2H-pyrido[4,3,2-de]呔 The salt form of the -3(7H)-ketone is prepared by treating the compound with an acid under the given conditions. The salt form No. 1-18 was determined in Table 1 below for the treatment of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1 ,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 The -3( 7H )-keto acid is provided in the "Acid" column and the conditions used during the treatment are provided in the "Conditions" column.

(8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro -2H-pyrido[4,3,2-de]呔 The free base polymorph of -3(7H)-one is prepared from solutions prepared in different solvents. Characterization of the free base form Nos. 19-24 is summarized in Table 2 and in the paragraphs referred to in Table 2.

Microscopy and other standard laboratory measurements to characterize the solid form, melting temperature, solvation and/or hydration and other physiochemical properties of the salt form and free base polymorph (including but not limited to solubility, PKa and LogP).

Table 1: (8S, 9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9 -dihydro-2H-pyrido[4,3,2-de]呔 -3(7H)-keto salt form

Table 2: Preparation of (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8, 9-dihydro-2H-pyrido[4,3,2-de]呔 Solvent in the polymorphic form of -3(7H)-ketone

HCl salt 1: Microscopy: large crystalline form. The results based on microscopy, XRPD (Table 3), DSC (Table 4) and TGA (Table 5) indicate that the HCl salt is in a mixed form of crystalline and amorphous forms and may be solvated.

Table 3: X-ray powder diffraction: Selected peaks of 1 * *The peak with a relative intensity less than 20% is not reported

Table 4: Results of differential scanning calorimetry Inclined to 300.00 ° C at 10.00 ° C / min

Table 5: Thermogravimetric analysis results of 1 Inclined to 300.00 ° C at 10.00 ° C / min Starting temperature = 25.7 ° C

Mesylate Salt 3: Microscopy: a mixed form of crystalline and amorphous. Melting point: 179 to 184 ° C, determined by a melting point apparatus. The results based on microscopy, XRPD (Table 6), DSC (Table 7) and TGA (Table 8) indicate that this mesylate salt is in mixed form and may be solvated.

Table 6: X-ray powder diffraction: selected peaks of 3 * * The peak with a relative intensity less than 20% is not reported

Table 7: Results of Differential Scanning Calorimetry for 3 Inclined to 300.00 ° C at 10.00 ° C / min

Table 8: Thermogravimetric analysis results of 3 Inclined to 300.00 ° C at 10.00 ° C / min Starting temperature = 25.64 ° C

Tosylate 4 from methanol-acetonitrile : (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4 -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3 (7 H) - one (190 mg, 0.5 mmol) was dissolved at 50 deg.] C in MeOH (2 mL) and CH ₃ CN (2 mL) in. Was then added and dissolved in MeOH (2 mL) and CH ₃ CN (2 mL) of a mixture of TsOH (190 mg, 1 mmol) . After 30 seconds, a white solid precipitated from the solution and the solution was cooled to 25 °C. Stirring was continued at 25 ° C for approximately 2 h. White solid was collected by filtration, which was CH ₃ CN (2 mL), dried and at 45 ℃ and dried under vacuum for 3 days. Three formulations were made with yields of 47%, 56%, and 56%, respectively, and the results were similar when measured by microscopy, XRPD, DSC, and TGA (inclination at 10.00 ° C/min to 300.00 ° C). The results provided herein are from one of the formulations. Microscopy: crystalline. The results based on microscopy, XRPD (Table 9) and DSC (Table 10) indicate that the tosylate salt is substantially in a single crystal form.

Table 9: X-ray powder diffraction: selected peaks of 4 * * The peak with a relative intensity less than 8% is not reported

Table 10: Results of the differential scanning calorimetry method of 4 Inclined to 400.00 ° C at 10.00 ° C / min

Tosylate 5 from dichloromethane-acetonitrile : (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H at 40 ° C -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3 (7 H) - after one (120 mg, 0.316 mmol) was suspended in CH ₂ Cl ₂ (5 mL) and the CH ₃ CN (5 mL), and add TsOH (66 mg, 0.348 mmol) , was added, the solution clarify. After 5 seconds, a white solid precipitated from the solution, stirring was continued at 25 ° C for 1 h, and filtered to give a white crystalline solid which was washed with CH ₃ CN (2 mL) and dried at 45 ° C under vacuum for 3 days. . Microscopy: birefringence, crystalline. The results based on microscopy, XRPD (Table 11), DSC (Tables 12a and 3a) and TGA (Table 12b and Figure 3b) indicate that the tosylate salt is substantially in a single crystal form.

Table 11: X-ray diffraction: selected peaks of 5* * The peak with a relative intensity less than 10% is not reported

Table 12a: Results of the differential scanning calorimetry method of 5 Inclined to 400.00 ° C at 10.00 ° C / min

Table 12b: TGA results of 5 Inclined to 400.00 ° C at 10.00 ° C / min

Tosylate 6, from acetone : (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2 at 25 ° C ,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3 (7 H) - one (120 mg, 0.316 mmol) was suspended in acetone (15 mL) of. After heating to 60 ° C, the mixture became clear and the temperature was lowered to 45 °C. TsOH (70 mg, 0.35 mmol) was then added, and after 30 seconds, a white solid precipitated from a solution cooled to 25 °C. Stirring was continued for 1 h at 25 °C. The white crystalline solid was collected by filtration, which was washed with acetone (8 mL) and dried at 45 ° C under vacuum for 3 days. Microscopy: birefringence, crystalline. The results based on microscopy, XRPD (Table 13), DSC (Tables 14a and 4a) and TGA (Table 14b and Figure 4b) indicate that the tosylate salt is substantially in a single crystal form.

Table 13: X-ray powder diffraction: selected peaks of 6 * * The peak with a relative intensity less than 10% is not reported

Table 14a: Results of differential scanning calorimetry of 6 Inclined to 400.00 ° C at 10.00 ° C / min

Table 14b: TGA results for 6 Inclined to 400.00 ° C at 10.00 ° C / min

Tosylate 7, from THF : (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2 at 25 ° C ,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3 (7 H) - one (120 mg, 0.316 mmol) was suspended in THF (6 mL) in. After heating to reflux, the mixture became clear and the temperature was lowered to 45 °C. TsOH (66 mg, 0.35 mmol) was then added, and after 1.5 min, a white solid precipitated from a solution cooled to 25 °C. Stirring was continued for 30 min at 25 °C. White crystalline solid was collected by filtration, the solid was CH ₂ Cl ₂ (10 mL) and washed with, and at 45 ℃ and vacuum dried for 3 days. Microscopy: birefringence, crystalline. The results based on microscopy, XRPD (Table 15), DSC (Tables 16a and 5a), and TGA (Table 16b and Figure 5b) indicate that the tosylate salt is substantially in a single crystal form.

Table 15: X-ray powder diffraction: selected peaks of 7* * The peak with a relative intensity less than 10% is not reported

Table 16a: Results of the differential scanning calorimetry method of 7 Inclined to 400.00 ° C at 10.00 ° C / min

Table 16b: TGA results of 7 Inclined to 400.00 ° C at 10.00 ° C / min

Tosylate 8, from acetone-THF: (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1 at 25 ° C ,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3( 7H )-one (400 mg, 1.05 mmol) was suspended in a mixture of acetone (27 mL) and THF (13 mL). Then TsOH (220 mg, 1.16 mmol) was added. After 30 seconds, a white solid precipitated from the solution, stirring was continued at 25 ° C for 30 min, and filtered to give a white crystalline solid, which was taken from acetone (10 mL) A mixture of 4-dioxane (4 mL) was washed and dried at 45 ° C under vacuum for 3 days. Microscopy: birefringence, crystalline. The results based on microscopy, XRPD (Tables 17 and 8), DSC (Tables 18a and 2a) and TGA (Table 18b and Figure 2b) indicate that the tosylate salt is substantially in a single crystal form. DVA was applied to this polymorph and showed a weight gain of 0.1574% from 0% to 95% RH (Figure 8).

Table 17: X-ray powder diffraction: selected peaks of 8* * The peak with a relative intensity less than 2% is not reported

Table 18a: Results of the differential scanning calorimetry method of 8 Inclined to 400.00 ° C at 10.00 ° C / min

Table 18b: TGA results of 8 Inclined to 400.00 ° C at 10.00 ° C / min

The XRPD peaks from the formulations of crystalline tosylate forms 4, 5, 6, 7, and 8 were similar. The superposition of the XRPD patterns of 5, 6, 7, and 8 is shown in FIG. The tosylate forms 4, 5, 6, 7, and 8 each have a single endothermic peak above 300 ° C as determined by DSC, while the other salts and free base forms have endotherms well below 300 ° C. These results indicate that (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5-yl )-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 The tosylate salt of -3( 7H )-ketone has a relatively stable single polymorph.

Isethionates 11, 12 and 13, from THF: Microscopy: crystalline. The DSC results between 11, 12 and 13 are inconsistent, but all three formulations have at least 2, and sometimes 3 endothermic peaks, including a broad endothermic peak below 150 ° C and a sharp endothermic peak around 272 ° C to 282 ° C. Unlike the above tosylate salt (whose TGA does not show any weight change below 200 ° C), the isethionate 11 has a weight loss of 5.72% at 134.8 ° C and the isethionate 12 at 139.06 ° C The weight loss of 7.034% is shown below. The results obtained for the isethionate indicate that this salt form exists in more than one form.

Phosphates 16, 17 and 18 : The microscopy, XRPD and DSC results were consistent with the crystalline and amorphous blends present in each of the three phosphate formulations.

Free base 19 from acetone : microscopy, XRPD and DSC results were consistent with the mixture of forms.

Table 19: X-ray powder diffraction: selected peaks of free base 19* * The peak with a relative intensity less than 30% is not reported

Table 20: Results of differential scanning calorimetry of free base 19 Inclined to 300.00 ° C at 10.00 ° C / min

Table 21: Results of thermogravimetric analysis of free base 19 Inclined to 300.00 ° C at 10.00 ° C / min Starting temperature = 25.64 ° C

Free base 20 from MeOH/EtOAc: Microscopy showed a birefringent crystalline form; the XRPD and DSC results were consistent with the mixture of forms.

Table 22: X-ray powder diffraction: selected peaks of free base 20 * * The peak with a relative intensity less than 14% is not reported

Table 23: Results of differential scanning calorimetry of free base 20 Inclined to 300.00 ° C at 10.00 ° C / min

Table 24: Thermogravimetric analysis results of free base 20 Inclined to 300.00 ° C at 10.00 ° C / min Starting temperature = 25.64 ° C

Free base form 21 from acetonitrile: a mixture of crystalline and amorphous forms was observed. DSC: The maximum value of the broad endothermic peak is close to 133.05 ° C, the maximum exothermic peak is close to 209.21 ° C and the maximum value of the peak endothermic peak is at 255.85 ° C. TGA: The weight loss was 3.7% at 142.46 °C and 4.259% at 256.48 °C. The peak of the peak resolved by XRPD and having a relative intensity greater than 19% ( The values of d are as follows: 16.58486, 11.49904, 6.33021, and 6.01178.

Free base form 22, from methanol: A mixture of crystalline and amorphous forms was observed which is similar in nature to the free base form 20.

Free base form 23, from isopropanol: The material produced is amorphous.

Free base form 24 from DMS/ethanol: A mixture of crystalline and amorphous forms was observed. DSC: endothermic peak: 52.20 ° C _max (12.54 J/g), 202.63 ° C _max (120.9 J/g), 249.34 ° C _max (40.65 J/g). TGA: ambient temperature to 120 ° C, weight loss of 0.9388%; weight loss of 17.47% at 223.8 ° C.

These results confirm that (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4- can be prepared from certain solvents Triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 The crystalline form of the -3( 7H )-ketone free base, however, was found to be not a single crystal form which is comparable in stability to the crystalline tosylate salt form provided herein.

(8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro -2H-pyrido[4,3,2-de]呔 Illustrative preparation of -3(7H)-ketotosylate

The following is an example of (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4 suitable for GMP mass production in a single crystal form. -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 A method of -3( 7H )-ketomonotosylate.

Variant 1: (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4 at 50 ° C to 54 ° C -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 A solution of TsOH (1.05 eq.) in THF (5 vol) was slowly added to a solution of -3( 7H )-one (12.4 g) in THF (40 vol). The mixture was stirred at this temperature for a further 30 minutes. The mixture was then concentrated to 3 to 5 vol by distillation at 30 ° C to 40 ° C and reduced pressure (vacuum: -0.07 MPa to -0.08 MPa). Further removal of the THF solvent was achieved by adding acetone (20 vol) and then distilling to 3 to 5 vol under reduced pressure and repeating three times. The mixture was cooled to 5 ° C and filtered under nitrogen. The solid was dried at 60 ° C for 17 hours. The title compound (16.0 g, 89% yield) was obtained as white crystals. LC-MS (ESI) m / z: 381 (M + 1) +. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 2.29 (s, 3H), 3.67 (s, 3H), 4.97-5.06 (m, 2H), 6.91-6.94 (dd, J ¹ = 2, J ² = 10.8 Hz, 1H), 7.06-7.19 (m, 5H), 7.19-7.51 (m, 4H), 7.74 (s, 1H), 7.87 (s, 1H), 10.32 (brs, 1H), 12.36 (s, 1H).

Variant 2: (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4 at 50 ° C to 54 ° C -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2- de ]呔 -3 (7 H) - one (0.24 kg) of THF (40 vol) was slowly added TsOH (1.05 eq.) Of THF (5 vol) was added. The mixture was stirred at this temperature for a further 30 minutes. The mixture was then concentrated to 3 to 5 vol by distillation at 30 ° C to 40 ° C and reduced pressure (vacuum: -0.07 MPa to -0.08 MPa). Further removal of the THF solvent was achieved by adding acetone (20 vol) and then distilling to 3 to 5 vol under reduced pressure and repeating three times. After removing the THF solvent and distilling, the mixture was reslurried with 12 vol of acetone at 50 ° C to 54 ° C for 14 to 15 hours. The mixture was then cooled to 5 ° C and filtered under nitrogen. The solid was dried at 60 ° C for 17 hours. The title compound (0.31 kg, 91.7% yield, 99.65% purity) was obtained as white crystals.

When ¹ H-NMR was carried out, a one-to-one integral ratio of the N-methyl group at 3.67 ppm from the free base to the methyl group at 2.29 ppm from toluenesulfonic acid indicated the formation of monotosylate.

When DVS was carried out at 25 ° C, the tosylate salt produced using the procedure described in Variation 2 showed that the weight increase (less than 1%) of the tosylate salt was negligible as the humidity increased from 0% to 95%. Excluding this, this indicates that the tosylate is not hygroscopic. (See Table 29).

The XRPD pattern in Figure 6 was collected using a PANalytical X'Pert PRO MPD PW3040 diffractometer that uses the incident beam of Cu radiation generated by the Optix telephoto trimming source. Use an elliptical gradient multilayer mirror to make CuKα X-rays (1.54059 ) is focused through the sample onto the detector. Prior to analysis, a niobium sample (NIST SRM 640d) was analyzed to verify that the observed Si 111 peak position coincided with the NIST certified position. Samples of the samples were sandwiched between 3 μm thick films and analyzed for transmission geometry. Use beam baffles, short anti-scatter extensions, and anti-scattering blades to minimize the background of air generation. A Soller slit (0.02 x 0.02 radians) is used for the incident and diffracted beams to minimize axial divergence. The diffraction pattern was collected using a scanning position sensitivity detector (X'Celerator) and a data collector software version 2.2b located 240 mm from the sample. XRPD data acquisition parameters: transmission mode, 45 kV and 40 mA X-ray tube setting, 1.00-39.99 °2θ scanning range, 0.017 °2θ step size, 1939 second collection time, 1.2 °/min scan rate, ° Divergence slit and 1.0 second sample rotation time. The entire list of peaks identified in the XRPD pattern or a subset thereof may be sufficient to characterize the polymorph obtained. The XRPD results provided (Table 25) are the results of the tosylate salt prepared using the procedure described in Variation 2. The data in Table 25 is derived from the XRPD spectrum provided in Figure 6.

DSC analysis was carried out on the tosylate salt prepared using the above variant 1 and variant 2 and similar results were observed.

A Mettler Toledo Differential Scanning Calorimeter 1 was used to obtain a DSC thermogram of the tosylate salt prepared using the procedure described in Variation 2. The temperature was ramped from 25 ° C to 400 ° C at 10 ° C / min. In the DSC diagram, an exothermic event is drawn in the upward direction. The data in Table 26 is derived from the DSC plot provided in Figure 7a. The thermal analysis shows a sharp onset of melting and the exotherm after the completion of the melting shows that melting occurs with decomposition. A flat baseline before reaching the onset of melting showed no thermal conversion prior to melting, indicating that the solvate formation did not change from one crystalline form to another.

A Mettler Toledo Thermogravimetric Analyzer/Differential Scanning Calorimeter 1 was used to obtain a TGA thermogram of the tosylate salt prepared using the procedure described in Variation 2. The temperature was ramped from 25 ° C to 400 ° C at 10 ° C / min. The TGA map is provided in Figure 7b. The trace shows a stable baseline until melting and decomposition begins. This shows that there is very little, if any, residual solvent or adsorbed water in the crystalline product, and the product is thermally stable until it reaches the melting point and begins to decompose.

The solid state ¹³ C cross-polarization magic angle rotation (CP/MAS) NMR spectrum of the tosylate salt prepared using the procedure described in Variation 2 was at 25 ° C in a Varian ^UNITY INOVA- 400 spectrometer (Larmor frequency (Larmor frequency) Frequency): Obtained on ¹³ C=100.543 MHz, ¹ H=399.787 MHz). The sample was loaded into a 4 mm PENCIL type zirconia rotor and rotated at 12 kHz and magic angle. The spectra were acquired with phase-modulated SPINAL-64 high-power ¹ H decoupling during the acquisition time, using a ¹ H pulse width of 2.6 μs (90°), a 5 ms ramp-up amplitude cross-polarization contact time, and a 30 ms acquisition time. , 20 second scan delay, approximately 45 kHz spectral width and 2799 data points and 400 co-added scans. Free induction decay (FID) was processed using a Varian/Agilent VNMR 6.1C software with an index line broadening factor of 65536 points and 10 Hz to improve the signal to noise ratio. The first three data points of the FID were backtested using a VNMR linear prediction algorithm to produce a flat baseline. The chemical shift of the spectral peak is based on the carbonyl carbon resonance of glycine at 176.5 ppm. The entire list of peaks identified in the ¹³ C NMR spectrum or a subset thereof may be sufficient to characterize the polymorph obtained. The data in Table 28 is derived from the spectra provided in Figure 12.

Table 25: XRPD: Selected peaks of tosylate prepared according to a large scale method* * The peak with a relative intensity less than 2% is not reported

Table 26: DSC results for tosylate prepared according to a large scale method Inclined from 25.00 ° C to 400.00 ° C at 10.00 ° C / min

Table 28: Solid state of tosylate prepared according to a large scale process ¹³ C NMR results

Table 29: DVS isotherm results for tosylate prepared according to a large scale method

Biological instance Example 1: Single agent cytotoxicity analysis in a mantle cell lymphoma cell line

Culture of the mantle cell lymphoma cell line Granta in Dulbecco's MEM (4.5 g/L glucose) containing 10% FBS, 2 mM L-glutamic acid, 100 U/mL penicillin (Penicillin) and 100 μg/mL streptomycin -519. The mantle cell lymphoma cell line Jeko-1 was cultured in RPMI 1640 containing 10% FBS, 100 U/mL penicillin and 100 μg/mL streptomycin. The mantle cell lymphoma cell line Z138 was cultured in IMDM containing 10% horse serum, 100 U/mL penicillin and 100 μg/mL streptomycin. All cell lines were maintained in a 37 ° C incubator with 5% CO ₂ until analysis. Granta-519, Jeko-1 and Z138 cells were seeded in 96-well plates at 1000 cells/well or 5000 cells/well. The cells were incubated overnight at 37 ° C and then treated with their respective growth media containing the free base in 0.1% DMSO at various concentrations ranging from 1000 nM to 0.32 nM. 0.1% DMSO was used as a mock treatment or control. After 4 days of incubation, cell survival was measured by CellTiter Glo (Promega) and the cell survival ratio was calculated relative to the control. Using GraphPad Prism5 drawing data and software to calculate the ₅₀ value IC. See Figures 10a and 10b.

Table 28: IC ₅₀ (nM) for 4 days analysis

Example 2: Single agent cytotoxicity analysis in head and neck tumor cell lines

Oral squamous cell carcinoma HSC-3 and HSC-4 cells were cultured in MEM + 10% FBS. Oral squamous cell carcinoma CAL 27 was cultured in DMEM + 10% FBS. The bottom cell-like squamous cell line PE/CA-PJ34 (pure line 12) was cultured in IMDM + 10% FBS. All cell lines were maintained in their respective growth medium containing 100 U/mL penicillin and 100 μg/mL streptomycin in a 37 ° C incubator with 5% CO ₂ until analysis. Depending on the growth rate of individual cell lines, cells were seeded in 96-well plates at different densities as shown in each figure at the time of analysis. The cells were incubated overnight at 37 ° C and then treated with their respective growth media containing the free base in 0.1% DMSO at various concentrations ranging from 5000 nM to 0.064 nM. 0.1% DMSO was used as a mock treatment or control. The medium and compound were supplemented every 5 days. After 7, 10, 11 or 13 days of incubation (as shown in each figure), cell viability was measured by CellTiter Glo (Promega) and cell viability ratio was calculated relative to control. Using GraphPad Prism5 drawing data and software to calculate the ₅₀ value IC. See Figures 11a and 11b.

Table 29: IC ₅₀ (nM)

Modifications and variations of the subject matter described in the above illustrative examples are contemplated by those skilled in the art. Any of the claimed inventions should be limited only as described in the accompanying claims.

All publications, including patents, patent applications, and published patent applications, which are hereby incorporated by reference in their entireties in the entireties in

Figure 1 provides a crystalline form of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl )-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 5 superposed one example tosylate of powder X-ray powder diffraction (XRPD) pattern of - -3 (7 H).

Figure 2a provides a crystalline form (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4- prepared from acetone-THF Triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 Differential Scanning Calorimetry (DSC) plot of -3( 7H )-ketotosylate polymorph 8.

Figure 2b provides a crystalline form (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4- from acetone-THF prepared. Triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 Thermogravimetric analysis (TGA) pattern of -3( 7H )-ketotosylate polymorph 8.

Figure 3a provides a crystalline form of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4- from DCM-ACN. Triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 DSC chart of -3( 7H )-ketotosylate polymorph 5.

Figure 3b provides a crystalline form of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4- from DCM-ACN Triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 TGA pattern of -3( 7H )-ketotosylate polymorph 5 .

Figure 4a provides a crystalline (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole prepared from acetone -5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 DSC chart of -3( 7H )-ketotosylate polymorph 6 .

Figure 4b provides a crystalline (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole prepared from acetone -5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 TGA pattern of -3( 7H )-ketotosylate polymorph 6 .

Figure 5a provides a crystalline form of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole prepared from THF -5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 DSC chart of -3( 7H )-ketotosylate polymorph 7.

Figure 5b provides a crystalline (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole prepared from THF -5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 TGA pattern of -3( 7H )-ketotosylate polymorph 7.

Figure 6 provides crystalline (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8 ,9-Dihydro-2H-pyrido[4,3,2-de]呔 X-ray powder diffraction (XRPD) spectrum of the -3 (7H)-ketotosylate polymorph.

Figure 7a provides a crystalline (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5-yl group )-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 DSC chart of the polymorphic form of -3( 7H )-ketotosylate.

Figure 7b provides a crystalline (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5-yl group )-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 TGA pattern of the polymorphic form of -3( 7H )-ketotosylate.

Figure 8 is a graph showing the crystalline form of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4 from acetone-THF. -triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 (7 H) - one polymorph tosylate X-ray powder diffraction (XRPD) 8 of the spectrum.

Figure 9 provides a crystalline form of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4- from acetone-THF. Triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 Dynamic vapor adsorption isotherm plot of -3( 7H )-ketotosylate polymorph 8.

Figure 10a provides data for a single agent cytotoxicity assay for free base in three mantle cell lymphoma cell lines (1000 cells/well).

Figure 10b provides data for a single agent cytotoxicity assay of free base in three mantle cell lymphoma cell lines (5000 cells/well).

Figure 11a provides data for a single agent cytotoxicity assay for free base in two head and neck cancer cell lines.

Figure 11b provides data for a single agent cytotoxicity assay for free base in a PE/CA-PJ34 head and neck cancer cell line.

Figure 12 provides (8S,9R)-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9 -dihydro-2H-pyrido[4,3,2-de]呔 Solid state ¹³ C NMR spectrum of the -3 (7H)-ketotosylate polymorph.

(no component symbol description)

Claims (34)

(8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8, 9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate. (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl as claimed in claim 1 )-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketotosylate, wherein the salt is in a substantially pure crystalline form. (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 as claimed in claim 1 or 2 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 (7 H) - -one tosylate, in which the at least one crystalline form of the salt type exhibits the following: a peak at 143.2,136.0,131.8,123.9,112.2,105.2 and 100.3 ppm ± 0.2 ppm of the Solid state ¹³ C NMR spectrum; differential scanning calorimetry thermogram with maximum endothermic between about 320 ° C and about 335 ° C; thermogravimetric thermogram indicating unsolvated material; from 0 to 95% Dynamic vapor adsorption isotherm diagram showing no significant weight change at relative humidity; including d values of approximately 11.9, 5.9, 4.9, 4.4, 4.3, 3.9, and 3.7 ( An X-ray powder diffraction pattern representing a characteristic peak; and an X-ray powder diffraction pattern including peaks at 2θ angles of 7.4, 15.1, 18.1, 20.1, 20.4, 22.6, and 24.0 ± 0.2 2θ angles. (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 as claimed in claim 1 or 2 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 (7 H) - -one tosylate, in which the at least one crystalline form of the salt type exhibits the following: a peak at 143.2,136.0,131.8,123.9,112.2,105.2 and 100.3 ppm ± 0.2 ppm of the Solid state ¹³ C NMR spectrum; including d values of about 11.9, 5.9, 4.9, 4.4, 4.3, 3.9, and 3.7 ( An X-ray powder diffraction pattern representing a characteristic peak; and an X-ray powder diffraction pattern including peaks at 2θ angles of 7.4, 15.1, 18.1, 20.1, 20.4, 22.6, and 24.0 ± 0.2 2θ angles. (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 as claimed in claim 1 or 2 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 (7 H) - -one tosylate, wherein the salt-based presentation includes a d value of about 3.7 and 11.9,5.9,4.9,4.4,4.3,3.9 ( a crystalline form of the X-ray powder diffraction pattern representing the characteristic peak. (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 as claimed in claim 1 or 2 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 (7 H) - -one tosylate, wherein the salt-based presentation includes a representation of at 7.4,15.1,18.1,20.1,20.4,22.6 and 24.0 ± 2θ angles of 0.2 2θ angle characteristic X-ray powder diffraction peaks The crystalline form of the pattern. (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 as claimed in claim 1 or 2 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3 (7 H) - -one tosylate, wherein the salt-based show in 143.2,136.0,131.8,123.9,112.2,105.2 and 100.3 ppm ± 0.2 ppm of the crystalline form having a peak of the solid-state ¹³ C NMR spectrum. (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4- as claimed in any one of claims 1 to 7 Triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-ketotosylate, wherein the crystalline form has a purity of at least about 99.5%. The tosylate salt according to any one of claims 1 to 8, wherein the crystalline form is substantially free of (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-( 1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 An amorphous form of -3( 7H )-one or a salt or solvate thereof. Preparation of (8 S, 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8 ,9-Dihydro-2 H -pyrido[4,3,2-de]呔 A method of crystallizing a form of -3( 7H )-ketotosylate, comprising the step (1): at elevated temperature, one or more independently selected from the group consisting of THF, acetone, methanol, acetonitrile, and DCM Step 1 In the presence of a solvent, (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3( 7H )-one is contacted with p-toluenesulfonic acid; step (2): allowing it to stand under conditions sufficient to precipitate the crystalline form; and step (3): isolating the crystalline form. The method of claim 10, wherein the elevated temperature is from about 30 ° C to about 70 ° C. The method of claim 10 or 11, wherein the one or more step 1 solvents are independently selected from the group consisting of methanol and acetonitrile. The method of claim 10 or 11, wherein the one or more step 1 solvents are independently selected from the group consisting of DCM and acetonitrile. The method of claim 10 or 11, wherein the one or more step 1 solvents are independently selected from the group consisting of acetone and THF. The method of claim 10, 11 or 14, wherein the solvent of step 1 is acetone. The method of claim 10, 11 or 14, wherein the solvent of step 1 is THF. The method of any one of claims 10 to 15, wherein the conditions sufficient to precipitate the crystalline form comprise cooling. The method of claim 17, wherein the conditions sufficient to precipitate the crystalline form comprise cooling to a temperature of 25 ° C or lower. The method of any one of clauses 10 to 18, further comprising the step (a) of: And the 4-fluorobenzaldehyde is contacted in a mixture comprising one or more steps (a) a solvent and titanium (III) chloride to prepare a first intermediate; and step (b): separating the first by separating the palms An enantiomer of the intermediate; and step (c): obtained by contacting the separated enantiomer of the first intermediate with a monohydrate hydrazine in one or more of the solvents of step (c) (8 S ,9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9 -dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketone. The method of claim 19, wherein the separate enantiomer of the first intermediate is contacted with a monohydrate hydrazine in one or more solvents selected from the group consisting of methanol, ethanol and acetonitrile in step (c). The method of claim 19 or 20, wherein the one or more steps (a) are independently selected from the group consisting of THF and methanol. The method of any one of clauses 10 to 18, further comprising the step (x): Contact with monohydrate hydrazine in one or more step x solvents to produce 5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazole-5 -yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketone; and step (y): separating (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-A by separation of palms Base-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketone. The method of claim 22, wherein the one or more steps (x) are independently selected from the group consisting of methanol, ethanol, and acetonitrile. (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1 prepared according to the method of any one of claims 10 to 23 ,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate. The tosylate salt of claim 24, wherein the salt exhibits a d value of about 7.4, 15.1, 18.1, 20.1, 20.4, 22.6, and 24.0 ( a crystalline form of the X-ray powder diffraction pattern representing the characteristic peak. The tosylate salt of claim 24 or 25, wherein the salt exhibits a single endothermic peak between room temperature and about 350 ° C in differential scanning calorimetry, wherein the single endothermic peak maximum occurs at about 320 °C to about 335 °C. The tosylate salt according to any one of claims 24 to 26, wherein the salt exhibits a thermal weight loss of less than 2% at a temperature of about 280 ° C or lower by thermogravimetric analysis. The tosylate salt of any one of claims 24 to 27, wherein the salt exhibits a hysteresis value of less than about 1% from RH 0% to RH 95% at about 25 ° C in dynamic vapor adsorption. A pharmaceutical composition comprising the tosylate salt of any one of claims 1 to 8 and 24 to 28 and a pharmaceutically acceptable excipient. The pharmaceutical composition of claim 29, wherein the composition is formulated for oral administration to an individual. (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8, 9-dihydro-2 H -pyrido[4,3,2-de]呔 Use of a -3( 7H )-ketotoluenesulfonate for the manufacture of a medicament for the treatment of cancer or a symptom thereof. The use of claim 31, wherein the cancer is bladder cancer, breast cancer, cervical cancer, colon cancer, colorectal cancer, Burkitt's lymphoma, nasopharyngeal carcinoma, EBV+gastric cancer, endometrial cancer, gastrointestinal Matrix tumor, glioblastoma, head and neck cancer, hepatocellular carcinoma, kidney cancer, leukemia, lung cancer, lymphoma, blastocytoma, melanoma, meningococcal tumor, ovarian cancer, pancreatic cancer, prostate cancer, kidney Cancer, small cell lung cancer, thyroid cancer or uterine cancer. The use of claim 31 or 32, wherein the individual is administered (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl) from about 0.3 μg/kg body weight to about 3.0 μg/kg body weight per day to the individual. 9-(1-methyl-1 H -1,2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 -3(7 H )-ketotosylate. The use of any one of claims 31 to 33, wherein the (8 S , 9 R )-5-fluoro-8-(4-fluorophenyl)-9-(1-methyl-1 H -1, 2,4-triazol-5-yl)-8,9-dihydro-2 H -pyrido[4,3,2-de]呔 The -3( 7H )-ketotoluenesulfonate is a crystalline form of any one of claims 3 to 9.